@article {2396, title = {Binding and Uptake of RGD-Containing Ligands to Cellular alpha(v)beta(3) Integrins}, journal = {International Journal of Peptide Research and Therapeutics}, volume = {15}, number = {1}, year = {2009}, note = {ISI Document Delivery No.: 430WTTimes Cited: 2Cited Reference Count: 34Cressman, Sonya Sun, Ying Maxwell, E. Jane Fang, Ning Chen, David D. Y. Cullis, Pieter R.}, month = {Mar}, pages = {49-59}, type = {Article}, abstract = {The cyclic peptide, cRGDf[N(me)]V, binds to the alpha(v)beta(3) integrin and can disrupt binding of the integrin to its natural ligands in the extracellular matrix. In this work, the ability of a water-soluble, fluorescently labeled variant of the RGD-containing peptide (cRGDfK-488) to bind to integrins on human umbilical vascular endothelial cells (HUVEC) and subsequently undergo endocytosis was characterized. This information was compared to the binding and uptake properties of an alpha(v)beta(3) integrin-specific monoclonal antibody, LM609X. The specificity of the RGD-containing peptide is assessed by comparison with control peptide that does not bind to the alpha(v)beta(3) integrin, cRADfK-488. Using a high purity construct, it is shown that the RGD ligand exhibits dissociation constants in the micromolar range whereas LM609X exhibits dissociation constants in the nanomolar range. However, the RGD ligand showed greater uptake following incubation at temperatures which permit endocytosis. A 7.4-fold increase in uptake of the RGD peptide was observed following a 1 h incubation with HUVEC at 37 degrees C (an endocytosis permissive temperature), as compared to that at 4 degrees C (an endocytosis prohibitive temperature). In contrast, only a 1.9-fold increase in cell-associated fluorescence was observed for similar incubations with LM609X. Results from fluorescence microscopy supports the notion that the RGD peptide is rapidly endocytosed at 37 degrees C as compared to LM609X. These results are discussed with regard to previous work indicating that RGD ligands enter cells by integrin-independent pathways. These studies provide well-controlled measures of how RGD ligands stimulate endocytosis. This may be of considerable interest for intracellular delivery of ligand-associated drugs in anti-angiogenic applications.}, keywords = {alpha(v)beta(3) integrins, ALPHA-V-BETA-3, ANGIOGENESIS, ANTAGONISTS, ANTITUMOR EFFICACY, CELLS, Endocytosis, EXPRESSION, IN-VIVO, PEPTIDES, RGD, TARGETED DRUG-DELIVERY, TUMOR VASCULATURE}, isbn = {1573-3149}, url = {://000265022900007}, author = {Cressman, S. and Sun, Y. and Maxwell, E. J. and Fang, N. and Chen, D. D. Y. and Cullis, P. R.} } @article {2663, title = {A Carboxy-Terminal Affinity Tag for the Purification and Mass Spectrometric Characterization of Integral Membrane Proteins}, journal = {Journal of Proteome Research}, volume = {8}, number = {5}, year = {2009}, note = {ISI Document Delivery No.: 441BWTimes Cited: 3Cited Reference Count: 22Wong, Julie P. Reboul, Emmanuelle Molday, Robert S. Kast, Juergen}, month = {May}, pages = {2388-2396}, type = {Article}, abstract = {G-protein-coupled receptors (GPCRs) and other structurally and functionally related membrane proteins represent particularly attractive targets for drug discovery. Integral membrane proteins are often difficult to purify from native contexts, and lack of sufficient quantities hampers subsequent structural and functional proteomic studies. We describe here an optimized enrichment strategy involving a membrane protein-compatible 1D4 affinity tag that is derived from the carboxy-terminal nine amino residues of bovine rhodopsin, and its corresponding tag-specific, high-affinity monoclonal antibody. When two GPCRs as well as two related ATP binding cassette (ABC) transporters are expressed in their functional forms in human cell lines, we have shown that a single detergent and wash condition can be employed for the purification of all said membrane proteins. Subsequent in-gel digestion with trypsin and mass spectrometric peptide analysis resulted in high sequence coverage for the ABC transporters ABCA1-1D4 and ABCA4-1D4. In contrast, digestion by various enzymatic combinations was necessary to obtain the best sequence coverage for affinity-enriched GPCRs CXCR4-1D4 and CCR5-1D4 as compared against other entries in an annotated spectrum library. Furthermore, specific enzyme combinations were necessary to produce suitable peptides for deducing N-glycosylation sites on CXCR4. Our results demonstrate that the 1D4-tag enrichment strategy is a versatile tool for the characterization of integral membrane proteins that can be employed for functional proteomic studies.}, keywords = {1D4 affinity tag, 2-DIMENSIONAL, annotated spectrum libraries, ATP binding cassette transporters, BINDING, ELECTROPHORESIS, EXPRESSION, G-protein-coupled, membrane protein enrichment, N-LINKED GLYCOSYLATION, PROTEOMICS, proteotypic peptides, RECEPTOR, RECEPTORS, RHODOPSIN, ROD OUTER SEGMENTS, SITES, SYNTHETIC PEPTIDES, Tandem mass spectrometry}, isbn = {1535-3893}, url = {://000265745300025}, author = {Wong, J. P. and Reboul, E. and Molday, R. S. and Kast, J.} } @article {2343, title = {Fruit-Surface Flavonoid Accumulation in Tomato Is Controlled by a SIMYB12-Regulated Transcriptional Network}, journal = {Plos Genetics}, volume = {5}, number = {12}, year = {2009}, note = {ISI Document Delivery No.: 542CTTimes Cited: 3Cited Reference Count: 58Adato, Avital Mandel, Tali Mintz-Oron, Shira Venger, Ilya Levy, Dorit Yativ, Merav Dominguez, Eva Wang, Zhonghua De Vos, Ric C. H. Jetter, Reinhard Schreiber, Lukas Heredia, Antonio Rogachev, Ilana Aharoni, Asaph}, month = {Dec}, pages = {23}, type = {Article}, abstract = {The cuticle covering plants{\textquoteright} aerial surfaces is a unique structure that plays a key role in organ development and protection against diverse stress conditions. A detailed analysis of the tomato colorless-peel y mutant was carried out in the framework of studying the outer surface of reproductive organs. The y mutant peel lacks the yellow flavonoid pigment naringenin chalcone, which has been suggested to influence the characteristics and function of the cuticular layer. Large-scale metabolic and transcript profiling revealed broad effects on both primary and secondary metabolism, related mostly to the biosynthesis of phenylpropanoids, particularly flavonoids. These were not restricted to the fruit or to a specific stage of its development and indicated that the y mutant phenotype is due to a mutation in a regulatory gene. Indeed, expression analyses specified three R2R3-MYB-type transcription factors that were significantly down-regulated in the y mutant fruit peel. One of these, SIMYB12, was mapped to the genomic region on tomato chromosome 1 previously shown to harbor the y mutation. Identification of an additional mutant allele that co-segregates with the colorless-peel trait, specific down-regulation of SlMYB12 and rescue of the y phenotype by overexpression of SlMYB12 on the mutant background, confirmed that a lesion in this regulator underlies the y phenotype. Hence, this work provides novel insight to the study of fleshy fruit cuticular structure and paves the way for the elucidation of the regulatory network that controls flavonoid accumulation in tomato fruit cuticle.}, keywords = {AFFECTS VEGETATIVE DEVELOPMENT, ARABIDOPSIS-THALIANA, EVOLUTION, EXPRESSION, GENE, IDENTIFICATION, LIQUID-CHROMATOGRAPHY, METABOLISM, PHENYLPROPANOID BIOSYNTHESIS, REGULATOR}, isbn = {1553-7390}, url = {://000273469700027}, author = {Adato, A. and Mandel, T. and Mintz-Oron, S. and Venger, I. and Levy, D. and Yativ, M. and Dominguez, E. and Wang, Z. H. and De Vos, R. C. H. and Jetter, R. and Schreiber, L. and Heredia, A. and Rogachev, I. and Aharoni, A.} } @article {2370, title = {Glucosamine conjugates bearing N,N,O-donors: potential imaging agents utilizing the [M(CO)(3)](+) core (M = Re, Tc)}, journal = {Dalton Transactions}, number = {42}, year = {2009}, note = {ISI Document Delivery No.: 508YBTimes Cited: 2Cited Reference Count: 32Bowen, Meryn L. Lim, Nathaniel C. Ewart, Charles B. Misri, Ripen Ferreira, Cara L. Haefeli, Urs Adam, Michael J. Orvig, Chris}, pages = {9216-9227}, type = {Article}, abstract = {The design rationale, synthesis and radiolabeling evaluation of four glucosamine conjugated ligands for the [Tc-99m(CO)(3)](+) core is described. The capability to bind the tricarbonyl core is initially demonstrated using the cold surrogate [Re(CO)(3)](+). The four compounds are competent chelates in binding [Tc-99m(CO)(3)](+) as labeling studies show, with yields ranging from 79 to 96\% and the resulting complexes showing stability in the presence of competing chelates for 24 h at 37 degrees C. The rhenium complexes were tested for hexokinase-catalysed phosphorylation, and the technetium complexes were tested for GLUT-1 mediated cell uptake - they showed a small amount of uptake but it was not glucose dependent, suggesting that it was not via the GLUT-1 transporters.}, keywords = {BIOMOLECULES, EXPRESSION, GLUT-1, HEXOKINASE, IN-VITRO, METAL-COMPLEXES, TC-99M, technetium, TRANSPORTERS, TRICARBONYL COMPLEXES}, isbn = {1477-9226}, url = {://000270973800020}, author = {Bowen, M. L. and Lim, N. C. and Ewart, C. B. and Misri, R. and Ferreira, C. L. and Hafeli, U. and Adam,Michael J. and Orvig, Chris} } @article {2370, title = {Glucosamine conjugates bearing N,N,O-donors: potential imaging agents utilizing the [M(CO)(3)](+) core (M = Re, Tc)}, journal = {Dalton Transactions}, number = {42}, year = {2009}, note = {ISI Document Delivery No.: 508YBTimes Cited: 2Cited Reference Count: 32Bowen, Meryn L. Lim, Nathaniel C. Ewart, Charles B. Misri, Ripen Ferreira, Cara L. Haefeli, Urs Adam, Michael J. Orvig, Chris}, pages = {9216-9227}, type = {Article}, abstract = {The design rationale, synthesis and radiolabeling evaluation of four glucosamine conjugated ligands for the [Tc-99m(CO)(3)](+) core is described. The capability to bind the tricarbonyl core is initially demonstrated using the cold surrogate [Re(CO)(3)](+). The four compounds are competent chelates in binding [Tc-99m(CO)(3)](+) as labeling studies show, with yields ranging from 79 to 96\% and the resulting complexes showing stability in the presence of competing chelates for 24 h at 37 degrees C. The rhenium complexes were tested for hexokinase-catalysed phosphorylation, and the technetium complexes were tested for GLUT-1 mediated cell uptake - they showed a small amount of uptake but it was not glucose dependent, suggesting that it was not via the GLUT-1 transporters.}, keywords = {BIOMOLECULES, EXPRESSION, GLUT-1, HEXOKINASE, IN-VITRO, METAL-COMPLEXES, TC-99M, technetium, TRANSPORTERS, TRICARBONYL COMPLEXES}, isbn = {1477-9226}, url = {://000270973800020}, author = {Bowen, M. L. and Lim, N. C. and Ewart, C. B. and Misri, R. and Ferreira, C. L. and Hafeli, U. and Adam,Michael J. and Orvig, Chris} } @article {2360, title = {Metabolic abnormalities in fronto-striatal-thalamic white matter tracts in schizophrenia}, journal = {Schizophrenia Research}, volume = {109}, number = {1-3}, year = {2009}, note = {ISI Document Delivery No.: 437VZTimes Cited: 3Cited Reference Count: 52Beasley, Clare L. Dwork, Andrew J. Rosoklija, Gorazd Mann, J. John Mancevski, Branislav Jakovski, Zlatko Davceva, Natasa Tait, Andrew R. Straus, Suzana K. Honer, William G.}, month = {Apr}, pages = {159-166}, type = {Article}, abstract = {{The anterior limb of the internal capsule (ALIC) is the major white matter tract providing reciprocal connections between the frontal cortex, striatum and thalamus. Mounting evidence suggests that this tract may be affected in schizophrenia, with brain imaging studies reporting reductions in white matter volume and density, changes in fractional anisotropy and reduced asymmetry. However, the molecular correlates of these deficits are currently unknown. The aim of this study was to identify alterations in protein and metabolite levels in the ALIC in schizophrenia. Samples were obtained post-mortem from individuals with schizophrenia (n = 15) and non-psychiatric controls (n = 13). Immunoreactivity for the myelin-associated protein myelin basic protein (MBP), and the axonal-associated proteins phosphorylated neurofilament and SNAP-25 was measured by enzyme-linked immunoadsorbent assay (ELISA). Metabolite concentrations were quantified by proton nuclear magnetic resonance (H-1 NMR) spectroscopy. Levels of myelin- or axonal-associated proteins did not differ between groups. Overall differences in metabolite concentrations were observed between the two groups (MANOVA F=2.685}, keywords = {ANTERIOR CINGULATE CORTEX, Axon, BASAL GANGLIA, composition, CORPUS-CALLOSUM, EXPRESSION, fiber, HUMAN INTERNAL CAPSULE, Internal capsule, LACTATE, MENTAL-ILLNESS, METABOLITE, myelin, POLARIZED-LIGHT, POSTMORTEM BRAINS, PREFRONTAL CORTEX, SEVERE}, isbn = {0920-9964}, url = {://000265516300023}, author = {Beasley, C. L. and Dwork, A. J. and Rosoklija, G. and Mann, J. J. and Mancevski, B. and Jakovski, Z. and Davceva, N. and Tait, A. R. and Straus, S. K. and Honer, W. G.} } @article {2376, title = {Plectosphaeroic Acids A, B, and C, Indoleamine 2,3-Dioxygenase Inhibitors Produced in Culture by a Marine Isolate of the Fungus Plectosphaerella cucumerina}, journal = {Organic Letters}, volume = {11}, number = {14}, year = {2009}, note = {ISI Document Delivery No.: 472NMTimes Cited: 1Cited Reference Count: 29Carr, Gavin Tay, Wendy Bottriell, Helen Andersen, Sarah K. Mauk, A. Grant Andersen, Raymond J.}, month = {Jul}, pages = {2996-2999}, type = {Article}, abstract = {Laboratory cultures of the fungus Plectosphaerella cucumerina obtained from marine sediments collected in Barkley Sound, British Columbia, yielded the novel alkaloids plectosphaeroic acids A (1) to C (3). The alkaloids 1-3 are inhibitors of indoleamine 2,3-dioxygenase (IDO).}, keywords = {CANCER, CELLS, chemotherapy, EXIGUAMINE-A, EXPRESSION, IDO, MECHANISM, METABOLITES, SUPPRESSION, TRYPTOPHAN DEGRADATION}, isbn = {1523-7060}, url = {://000268138800011}, author = {Carr, G. and Tay, W. and Bottriell, H. and Andersen, S. K. and Mauk, A. G. and Andersen, R. J.} } @article {2350, title = {Rationale, design and baseline characteristics of the PROJECT II study: PROpofol CardioproTECTion for Type II diabetics A randomized, controlled trial of high-dose propofol versus isoflurane preconditioning in patients undergoing on-pump coronary artery b}, journal = {Contemporary Clinical Trials}, volume = {30}, number = {4}, year = {2009}, note = {ISI Document Delivery No.: 456NPTimes Cited: 1Cited Reference Count: 23Ansley, David M. Raedschelders, Koen Chen, David D. Y. Choi, Peter T.}, month = {Jul}, pages = {380-385}, type = {Article}, abstract = {Diabetes mellitus is a leading cause of death globally and results in significant morbidity and mortality following surgery. After cardiac surgery, diabetic patients are especially at risk for low cardiac output syndrome, which can quadruple the risk for postoperative death. Attempts to prevent low cardiac output syndrome have focused on increasing myocardial tolerance to ischemia (preconditioning), which involves the myocardial mitochondrial ATP-regulated K-ATP channel. G-protein initiation, nitric oxide synthase, and protein kinase C. Unfortunately, the signal transduction pathways required for preconditioning are corrupted in diabetes. Effective antioxidant intervention during ischemia-reperfusion appears important for preserving myocardial function: thus, alleviating oxidant-mediated post-ischemic injury by increasing antioxidant defenses (cardioprotection) is an alternative to preconditioning. Our previous work suggests that propofol(2,6-diisopropylphenol), an intravenous anesthetic with antioxidant potential, may confer cardioprotection. In this paper, we describe the rationale and methodology of the Pro-TECT II Study, a Phase II randomized controlled trial designed to explore the relationships of biomarkers of oxidative or nitrosative stress in diabetes, to determine the effect of propofol cardioprotection to counteract these effects in patients undergoing elective primary coronary bypass graft surgery with cardiopulmonary bypass, and to provide feasibility and sample size data needed to conduct Phase III trials. (C) 2009 Elsevier Inc. All rights reserved.}, keywords = {15-F-2T-ISOPROSTANE FORMATION, antioxidant, apoptosis, CAPACITY, Cardiopulmonary bypass, Diabetes mellitus, EXPRESSION, F2-isoprostanes, HYPERGLYCEMIA, INJURY, MORTALITY, MYOCARDIAL-INFARCTION, Nitric oxide synthase type III, PREDICTORS, Propofol, SHORT-TERM}, isbn = {1551-7144}, url = {://000266853900014}, author = {Ansley, D. M. and Raedschelders, K. and Chen, D. D. Y. and Choi, P. T.} } @article {2298, title = {Biomimetic synthesis of the IDO inhibitors exiguamine A and B}, journal = {Nature Chemical Biology}, volume = {4}, number = {9}, year = {2008}, note = {ISI Document Delivery No.: 339SNTimes Cited: 7Cited Reference Count: 19Volgraf, Matthew Lumb, Jean-Philip Brastianos, Harry C. Carr, Gavin Chung, Marco K. W. Muenzel, Martin Mauk, A. Grant Andersen, Raymond J. Trauner, Dirk}, month = {Sep}, pages = {535-537}, type = {Article}, abstract = {Biomimetic synthesis is an attempt to assemble natural products along biosynthetic lines without recourse to the full enzymatic machinery of nature. We exemplify this with a total synthesis of exiguamine A and the newly isolated natural product exiguamine B. The most noteworthy feature of this work is an oxidative endgame drawing from the complex chemistry of catecholamines, which allows for ready access to a new class of nanomolar indoleamine-2,3-dioxygenase inhibitors.}, keywords = {3-DIOXYGENASE, ANALOGS, CANCER, CASCADE, CELLS, ELECTROCYCLIZATIONS, EXPRESSION, INDOLEAMINE 2}, isbn = {1552-4450}, url = {://000258597700011}, author = {Volgraf, M. and Lumb, J. P. and Brastianos, H. C. and Carr, G. and Chung, M. K. W. and Munzel, M. and Mauk, A. G. and Andersen, R. J. and Trauner, D.} } @article {2303, title = {The crystal structure of MexR from Pseudomonas aeruginosa in complex with its antirepressor ArmR}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {105}, number = {39}, year = {2008}, note = {ISI Document Delivery No.: 386WWTimes Cited: 15Cited Reference Count: 41Wilke, Mark S. Heller, Markus Creagh, A. Louise Haynes, Charles A. McIntosh, Lawrence P. Poole, Keith Strynadka, Natalie C. J.}, month = {Sep}, pages = {14832-14837}, type = {Article}, abstract = {The intrinsic antimicrobial resistance of the opportunistic human pathogen Pseudomonas aeruginosa is compounded in mutant strains that overexpress multidrug efflux pumps such as the prominent drug-proton antiporter, MexAB-OprM. The primary regulator of the mexAB-oprM operon is the MarR family repressor, MexR. An additional repressor, NalC, also regulates mexAB-oprM by controlling expression of ArmR, an antirepressor peptide that is hypothesized to prevent the binding of MexR to its cognate DNA operator via an allosteric protein-peptide interaction. To better understand how ArmR modulates MexR, we determined the MexR-binding region of ArmR as its C-terminal 25 residues and solved the crystal structure of MexR in a 2: 1 complex with this ArmR fragment at 1.8 angstrom resolution. This structure reveals that the C-terminal residues of ArmR form a kinked alpha-helix, which occupies a pseudo-symmetrical and largely hydrophobic binding cavity located at the centre of the MexR dimer. Although the ArmR-binding cavity partially overlaps with the small molecule effector-binding sites of other MarR family members, it possesses a larger and more complex binding surface to accommodate the greater size and specific physicochemical properties of a peptide effector. Comparison with the structure of apo-MexR reveals that ArmR stabilizes a dramatic conformational change that is incompatible with DNA-binding. Thus, this work defines the structural mechanism by which ArmR allosterically derepresses MexR-controlled gene expression in P. aeruginosa and reveals important insights into the regulation of multidrug resistance.}, keywords = {DEINOCOCCUS-RADIODURANS, DNA-BINDING, EXPRESSION, GENE, gene regulation, MarR, MARR FAMILY, MECHANISM, mexAB-oprM, MULTIDRUG EFFLUX OPERON, OPRM, PA3719, protein peptide, REPRESSOR, TRANSCRIPTIONAL REGULATOR HUCR}, isbn = {0027-8424}, url = {://000261914300004}, author = {Wilke, M. S. and Heller, M. and Creagh, A. L. and Haynes, C. A. and McIntosh, L. P. and Poole, K. and Strynadka, N. C. J.} } @article {2277, title = {Phosphorylation of U24 from Human Herpes Virus type 6 (HHV-6) and its potential role in mimicking myelin basic protein (MBP) in multiple sclerosis}, journal = {Febs Letters}, volume = {582}, number = {18}, year = {2008}, note = {ISI Document Delivery No.: 332UKTimes Cited: 2Cited Reference Count: 31Tait, Andrew R. Straus, Suzana K.}, month = {Aug}, pages = {2685-2688}, type = {Article}, abstract = {Myelin basic protein (MBP) from multiple sclerosis ( MS) patients contains lower levels of phosphorylation at Thr97 than normal individuals. The significance of phosphorylation at this site is not fully understood, but it is proposed to play a role in the normal functioning of MBP. Human Herpesvirus Type 6 encodes the protein U24, which has tentatively been implicated in the pathology of MS. U24 shares a 7 amino acid stretch encompassing the Thr97 phosphorylation site of MBP: PRTPPPS. We demonstrate using a combination of mass spectrometry, thin layer chromatography and autoradiography, that U24 can be phosphorylated at the equivalent threonine. Phospho-U24 may confound signalling or other pathways in which phosphorylated MBP may participate, precipitating a pathological process.}, keywords = {BINDING, EXPRESSION, HUMAN-HERPESVIRUS-6, IDENTIFICATION, KINASE, MEMBRANE MICRODOMAINS, mimicry, myelin, PHOSPHORYLATION, POSTTRANSLATIONAL MODIFICATIONS, SITE, T-ANTIGEN, U24}, isbn = {0014-5793}, url = {://000258108400002}, author = {Tait, A. R. and Straus, S. K.} } @article {2037, title = {Synthesis of indoleamine 2,3-dioxygenase inhibitory analogues of the sponge alkaloid exiguamine A}, journal = {Journal of Medicinal Chemistry}, volume = {51}, number = {9}, year = {2008}, note = {ISI Document Delivery No.: 295USTimes Cited: 17Cited Reference Count: 20Carr, Gavin Chung, Marco K. W. Mauk, A. Grant Andersen, Raymond J.}, month = {May}, pages = {2634-2637}, type = {Article}, abstract = {Synthetic analogues of the sponge natural product exiguamine A (3) have been prepared and evaluated for their ability to inhibit indoleamine 2,3-dioxygenase in vitro.}, keywords = {CANCER, CATABOLISM, CELLS, chemotherapy, EXPRESSION, IDO, IMMUNE TOLERANCE, MECHANISM, SMALL-MOLECULE INHIBITORS, TRYPTOPHAN DEGRADATION}, isbn = {0022-2623}, url = {://000255500000008}, author = {Carr, G. and Chung, M. K. W. and Mauk, A. G. and Andersen, R. J.} } @article {1518, title = {Exploring the mode-of-action of bioactive compounds by chemical-genetic profiling in yeast}, journal = {Cell}, volume = {126}, number = {3}, year = {2006}, note = {ISI Document Delivery No.: 075KITimes Cited: 111Cited Reference Count: 68Parsons, Ainslie B. Lopez, Andres Givoni, Inmar E. Williams, David E. Gray, Christopher A. Porter, Justin Chua, Gordon Sopko, Richelle Brost, Renee L. Ho, Cheuk-Hei Wang, Jiyi Ketela, Troy Brenner, Charles Brill, Julie A. Fernandez, G. Esteban Lorenz, Todd C. Payne, Grego S. Ishihara, Satoru Ohya, Yoshikazu Andrews, Brenda Hughes, Timothy R. Frey, Brendan J. Graham, Todd R. Andersen, Raymond J. Boone, Charles}, month = {Aug}, pages = {611-625}, type = {Article}, abstract = {Discovering target and off-target effects of specific compounds is critical to drug discovery and development. We generated a compendium of "chemical-genetic interaction" profiles by testing the collection of viable yeast haploid deletion mutants for hypersensitivity to 82 compounds and natural product extracts. To cluster compounds with a similar mode-of-action and to reveal insights into the cellular pathways and proteins affected, we applied both a hierarchical clustering and a factorgram method, which allows a gene or compound to be associated with more than one group. In particular, tamoxifen, a breast cancer therapeutic, was found to disrupt calcium homeostasis and phosphatidylserine (PS) was recognized as a target for papuamide B, a cytotoxic lipopeptide with anti-HIV activity. Further, the profile of crude extracts resembled that of its constituent purified natural product, enabling detailed classification of extract activity prior to purification. This compendium should serve as a valuable key for interpreting cellular effects of novel compounds with similar activities.}, keywords = {1, 3-BETA-D-GLUCAN SYNTHASE, BREAST-CANCER CELLS, DELETION MUTANTS, drug, EXPRESSION, GENOME-WIDE ANALYSIS, HAPLOINSUFFICIENCY, INDUCED, SACCHAROMYCES-CEREVISIAE, THEONELLA-SWINHOEI, TRANSCRIPTION FACTOR}, isbn = {0092-8674}, url = {://000239883400019}, author = {Parsons, A. B. and Lopez, A. and Givoni, I. E. and Williams, D. E. and Gray, C. A. and Porter, J. and Chua, G. and Sopko, R. and Brost, R. L. and Ho, C. H. and Wang, J. Y. and Ketela, T. and Brenner, C. and Brill, J. A. and Fernandez, G. E. and Lorenz, T. C. and Payne, G. S. and Ishihara, S. and Ohya, Y. and Andrews, B. and Hughes, T. R. and Frey, B. J. and Graham, T. R. and Andersen, R. J. and Boone, C.} } @article {1621, title = {The unfolding and folding dynamics of TNfnALL probed by single molecule force-ramp spectroscopy}, journal = {Polymer}, volume = {47}, number = {7}, year = {2006}, note = {ISI Document Delivery No.: 030JETimes Cited: 10Cited Reference Count: 53}, month = {Mar}, pages = {2548-2554}, type = {Article}, abstract = {Tenascin, an important extracellular matrix protein, is subject to stretching force under physiological conditions and plays important roles in regulating the cell-matrix interactions. Using the recently developed single molecule force-ramp spectroscopy, we investigated the unfolding-folding kinetics of a recombinant tenascin fragment TNfnALL. Our results showed that all the 15 FnIII domains in TNfnALL have similar spontaneous unfolding rate constant at zero force, but show great difference in their folding rate constants. Our results demonstrated that single molecule force-ramp spectroscopy is a powerful tool for accurate determination of the kinetic parameters that characterize the unfolding and folding reactions. We anticipate that single molecule force-ramp spectroscopy will become a versatile addition to the single molecule manipulation tool box and greatly expand the scope of single molecule force spectroscopy. (c) 2006 Elsevier Ltd. All rights reserved.}, keywords = {ADHESION, DOMAIN, EXPRESSION, III, IMMUNOGLOBULIN, MATRIX PROTEINS, MECHANICAL STABILITY, microscopy, MUSCLE PROTEIN TITIN, tenascin, UBIQUITIN}, isbn = {0032-3861}, url = {://000236629900037}, author = {Wang, M. J. and Cao, Y. and Li, H. B.} } @article {1149, title = {Quantitative proteomic analysis of sokotrasterol sulfate-stimulated primary human endothelial cells}, journal = {Molecular \& Cellular Proteomics}, volume = {4}, number = {2}, year = {2005}, note = {ISI Document Delivery No.: 902UFTimes Cited: 9Cited Reference Count: 40}, month = {Feb}, pages = {191-204}, type = {Article}, abstract = {The endothelium forms a continuous monolayer at the interface between blood and tissue and contributes significantly to the sensing and transducing of signals between blood and tissue. New blood vessel formation, or angiogenesis, is initiated by the activation of endothelial cells and is an important process required for various pathological and physiological situations. This study used cleavable isotope-coded affinity tag reagents combined with mass spectrometry to investigate the molecular basis of a recently discovered angiogenesis-promoting steroid, sokotrasterol sulfate. Changes in the relative abundances of over 1000 proteins within human endothelial cells treated with sokotrasterol sulfate and vehicle-treated cells were identified and quantitated using this technique. A method that examines the entire ensemble of quantitative measurements was developed to identify proteins that showed a statistically significant change in relative abundance resulting from treatment with sokotrasterol sulfate. A total of 93 proteins was significantly up-regulated, and 37 were down-regulated in response to sokotrasterol sulfate stimulation of endothelial cells. Among the up-regulated proteins, several were identified that are novel to endothelial cells and are likely involved in cell communication and morphogenesis. These findings are consistent with a role for sokotrasterol sulfate in endothelial sprouting.}, keywords = {CANCER, DIFFERENTIATION, EXPRESSION, GENE, GROWTH-FACTOR, ISCHEMIC-HEART-DISEASE, MASS-SPECTROMETRY, MOLECULE, THERAPEUTIC ANGIOGENESIS, VEGF}, isbn = {1535-9476}, url = {://000227381300008}, author = {Karsan, A. and Pollet, I. and Yu, L. R. and Chan, K. C. and Conrads, T. P. and Lucas, D. A. and Andersen, R. J. and Veenstra, T.} } @article {1301, title = {RS1, a discoidin domain-containing retinal cell adhesion protein associated with X-linked retinoschisis, exists as a novel disulfide-linked octamer}, journal = {Journal of Biological Chemistry}, volume = {280}, number = {11}, year = {2005}, note = {ISI Document Delivery No.: 905HQTimes Cited: 33Cited Reference Count: 25}, month = {Mar}, pages = {10721-10730}, type = {Article}, abstract = {RS1, also known as retinoschisin, is an extracellular protein that plays a crucial role in the cellular organization of the retina. Mutations in RS1 are responsible for X-linked retinoschisis, a common, early-onset macular degeneration in males that results in a splitting of the inner layers of the retina and severe loss in vision. RS1 is assembled and secreted from photoreceptors and bipolar cells as a homo-oligomeric protein complex. Each subunit consists of a 157-amino acid discoidin domain flanked by two small segments of 39 and 5 amino acids. To begin to understand how the structure of RS1 relates to its role in retinal cell adhesion and X-linked retinoschisis, we have determined the subunit organization and disulfide bonding pattern of RS1 by SDS gel electrophoresis, velocity sedimentation, and mass spectrometry. Our results indicate that RS1 exists as a novel octamer in which the eight subunits are joined together by Cys(59)-Cys(223) intermolecular disulfide bonds. Subunits within the octamer are further organized into dimers mediated by Cys(40)-Cys(40) bonds. These cysteines lie just outside the discoidin domain indicating that these flanking segments primarily function in the octamerization of RS1. Within the discoidin domain, two cysteine pairs (Cys(63)-Cys(219) and Cys(110)-Cys(142)) form intramolecular disulfide bonds that are important in protein folding, and one cysteine (Cys(83)) exists in its reduced state. Because mutations that disrupt subunit assembly cause X-linked retinoschisis, the assembly of RS1 into a disulfide-linked homo-octamer appears to be critical for its function as a retinal cell adhesion protein.}, keywords = {C2 DOMAIN, COAGULATION-FACTOR-V, CRYSTAL-STRUCTURES, EXPRESSION, FACTOR-VIII, GENE, JUVENILE RETINOSCHISIS, MUTATIONS, PHOTORECEPTOR, RESOLUTION}, isbn = {0021-9258}, url = {://000227559600118}, author = {Wu, W. W. H. and Wong, J. P. and Kast, J. and Molday, R. S.} } @article {1023, title = {The NeuC protein of Escherichia coli K1 is a UDP N-acetylglucosamine 2-epimerase}, journal = {Journal of Bacteriology}, volume = {186}, number = {3}, year = {2004}, note = {ISI Document Delivery No.: 766HUTimes Cited: 18Cited Reference Count: 43}, month = {Feb}, pages = {706-712}, type = {Article}, abstract = {The K1 capsule is an essential virulence determinant of Escherichia coli strains that cause meningitis in neonates. Biosynthesis and transport of the capsule, an alpha-2,8-linked polymer of sialic acid, are encoded by the 17-kb kps gene cluster. We deleted neuC, a K1 gene implicated in sialic acid synthesis, from the chromosome of EV36, a K-12-K1 hybrid, by allelic exchange. Exogenously added sialic acid restored capsule expression to the deletion strain (DeltaneuC), confirming that NeuC is necessary for sialic acid synthesis. The deduced amino acid sequence of NeuC showed similarities to those of UDP-N-acetylglucosamine (GlcNAc) 2-epimerases from both prokaryotes and eukaryotes. The NeuC homologue from serotype III Streptococcus agalactiae complements DeltaneuC. We cloned the neuC gene into an intein expression vector to facilitate purification. We demonstrated by paper chromatography that the purified neuC gene product catalyzed the formation of [2-C-14]acetamidoglucal and [N-C-14]acetylmannosamine (ManNAc) from UDP-[C-14]GlcNAc. The formation of reaction intermediate 2-acetamidoglucal with the concomitant release of UDP was confirmed by proton and phosphorus nuclear magnetic resonance spectroscopy. NeuC could not use GlcNAc as a substrate. These data suggest that neuC encodes an epimerase that catalyzes the formation of ManNAc from UDP-GlcNAc via a 2-acetamidoglucal intermediate. The unexpected release of the glucal intermediate and the extremely low rate of ManNAc formation likely were a result of the in vitro assay conditions, in which a key regulatory molecule or protein was absent.}, keywords = {2-EPIMERASE/N-ACETYLMANNOSAMINE KINASE, ACETYLNEURAMINIC ACID SYNTHETASE, EXPRESSION, GENE-PRODUCT, NEISSERIA-MENINGITIDIS, POLYSACCHARIDE BIOSYNTHESIS, POLYSIALIC ACID, RAT-LIVER, SEQUENCE, SIALIC-ACID}, isbn = {0021-9193}, url = {://000188371600014}, author = {Vann, W. F. and Daines, D. A. and Murkin, A. S. and Tanner, M. E. and Chaffin, D. O. and Rubens, C. E. and Vionnet, J. and Silver, R. P.} } @article {671, title = {HTI-286, a synthetic analogue of the tripeptide hemiasterlin, is a potent antimicrotubule agent that circumvents P-glycoprotein-mediated resistance in vitro and in vivo}, journal = {Cancer Research}, volume = {63}, number = {8}, year = {2003}, note = {ISI Document Delivery No.: 668TETimes Cited: 57Cited Reference Count: 32}, month = {Apr}, pages = {1838-1845}, type = {Article}, abstract = {Hemiasterlin is a natural product derived from marine sponges that, like other structurally diverse peptide-like molecules, binds to the Vincapeptide site in tubulin, disrupts normal microtubule dynamics, and, at stoichiometric amounts, depolymerizes microtubules. Total synthesis of hemiasterlin and its analogues has been accomplished, and optimal pharmacological features of the series have been explored. The biological profile of one analogue, HTI-286, was studied here. HTI-286 inhibited the polymerization of purified tubulin, disrupted microtubule organization in cells, and induced mitotic arrest, as well as apoptosis. HTI-286 was a potent inhibitor of proliferation (mean IC50 = 2.5 +/- 2.1 nm in 18 human tumor cell lines) and had substantially less interaction with multidrug resistance protein (P-glycoprotein) than currently used antimicrotubule agents, including paclitaxel, docetaxel, vinorelbine, or vinblastine. Resistance to HTI-286 was not detected in cells overexpressing the drug transporters MRP1 or MXR. In athymic mice implanted with human tumor xenografts, HTI-286 administered i.v. in saline inhibited the growth of numerous human tumors derived from carcinoma of the skin, breast, prostate, brain, and colon. Marked tumor regression was observed when used on established tumors that were >1 gram in size. Moreover, HTI-286 inhibited the growth of human tumor xenografts (e.g., HCT-15, DLD-1, MX-1W, and KB-8-5) where paclitaxel and vincristine were ineffective because of inherent or acquired resistance associated with P-glycoprotein. Efficacy was also achieved with p.o. administration of HTI-286. These data suggest that HTI-286 has excellent preclinical properties that may translate into superior clinical activity, as well as provide a useful synthetic reagent to probe the drug contact sites of peptide-like molecules that interact with tubulin.}, keywords = {ADRIAMYCIN, BETA-TUBULIN, CANCER, CARCINOMA-CELLS, CYTOTOXIC PEPTIDES, DOLASTATIN ANALOG, DRUG-RESISTANCE, EXPRESSION, MECHANISMS, MULTIDRUG-RESISTANCE}, isbn = {0008-5472}, url = {://000182308600021}, author = {Loganzo, F. and Discafani, C. M. and Annable, T. and Beyer, C. and Musto, S. and Hari, M. and Tan, X. Z. and Hardy, C. and Hernandez, R. and Baxter, M. and Singanallore, T. and Khafizova, G. and Poruchynsky, M. S. and Fojo, T. and Nieman, J. A. and Ayral-Kaloustian, S. and Zask, A. and Andersen, R. J. and Greenberger, L. M.} } @article {441, title = {Sequential anaerobic-aerobic treatment of soil contaminated with weathered aroclor 1260}, journal = {Environmental Science \& Technology}, volume = {36}, number = {1}, year = {2002}, note = {ISI Document Delivery No.: 509RCTimes Cited: 25Cited Reference Count: 36}, month = {Jan}, pages = {100-103}, type = {Article}, abstract = {Soil contaminated with weathered Aroclor 1260 was bioremediated by sequential anaerobic and aerobic laboratory-scale treatment The initial concentration was 59 mug of PCBs/g of soil. Following 4 months of anaerobic treatment with an enrichment culture, all of the major components in Aroclor 1260 were completely or partially transformed to less chlorinated PCB congeners. The major products of reductive dechlorination were 24-24-tetrachlorobiphenyl and 24-26-tetrachlorobiphenyl, and the average chlorine substituents per PCB molecule decreased from 6.4 to 5.2. The molar concentration of PCBs did not decrease during the anaerobic treatment. All of the major products formed during the anaerobic treatment were degraded in the subsequent aerobic treatment using Burkholderia sp. strain LB400. After 28 days of the aerobic treatment, the concentration of PCBs was reduced to 20 mug/g of soil. PCBs were not significantly removed in aerobic treatments unless they were bioaugmented with LB400. Also, PCB degradation was not detected in soil bioaugmented with LB400 without prior anaerobic treatment. These results confirm the potential for extensive biological destruction of highly chlorinated, weathered PCB congeners in soil.}, keywords = {BACTERIA, BIODEGRADATION, CLONING, ENCODING BIPHENYL, EXPRESSION, GENE-CLUSTER, PCBS, POLYCHLORINATED BIPHENYL-DEGRADATION, PSEUDOMONAS STRAIN LB400, REDUCTIVE DECHLORINATION}, isbn = {0013-936X}, url = {://000173162600031}, author = {Master, E. R. and Lai, V. W. M. and Kuipers, B. and Cullen, W. R. and Mohn, W. W.} } @article {4689, title = {Undercarboxylation of recombinant prothrombin revealed by analysis of gamma-carboxyglutamic acid using capillary electrophoresis and laser-induced fluorescence}, journal = {Febs Letters}, volume = {445}, number = {2-3}, year = {1999}, note = {ISI Document Delivery No.: 175YCTimes Cited: 6Cited Reference Count: 37}, month = {Feb}, pages = {256-260}, type = {Article}, abstract = {The gamma-carboxyglutamic acid (Gla) content of several variants of human prothrombin has been measured by using capillary electrophoresis and laser-induced fluorescence (CE-LIF). Both plasma-derived prothrombin and recombinant prothrombin contain ten residues of Gla per molecule of protein. In contrast, a variant of human prothrombin (containing the second kringle domain of bovine prothrombin) was separated into two populations that differed in their Gla content. Direct measurement of the Gla content showed an association with the presence or absence of the calcium-dependent conformational change that is required for prothombinase function. Thus, the CE-LIF assay is useful in determining the carboxylation status of recombinant proteins. (C) 1999 Federation of European Biochemical Societies.}, keywords = {baby hamster kidney, calcium, capillary electrophoresis, cell, EXPRESSION, FACTOR-X, gamma carboxy glutamic acid, GLUTAMIC-ACID, HUMAN PROTEIN-C, LASER-INDUCED FLUORESCENCE, METAL, PHOSPHOLIPID-BINDING-PROPERTIES, PROTHROMBIN, PURIFICATION, recombinant protein, RESIDUES, VITAMIN-K}, isbn = {0014-5793}, url = {://000079122800007}, author = {Vo, H. C. and Britz-McKibbin, P. and Chen, D. D. Y. and MacGillivray, R. T. A.} } @article {3708, title = {Interaction of soluble cellooligosaccharides with the N-terminal cellulose-binding domain of Cellulomonas fimi CenC .2. NMR and ultraviolet absorption spectroscopy}, journal = {Biochemistry}, volume = {35}, number = {44}, year = {1996}, note = {ISI Document Delivery No.: VR098Times Cited: 68Cited Reference Count: 46}, month = {Nov}, pages = {13895-13906}, type = {Article}, abstract = {The N-terminal cellulose-binding domain (CBDN1) from Cellulomonas fimi beta-1,4-glucanase CenC binds amorphous but not crystalline cellulose. To investigate the structural and thermodynamic bases of cellulose binding, NMR and difference ultraviolet absorbance spectroscopy were used in parallel with calorimetry (Tomme, P., Creagh, A. L., Kilburn, D. G., \& Haynes, C. A., (1996) Biochemistry 35, 13885-13894] to characterize the interaction of soluble cellooligosaccharides with CBDN1 Association constants, determined from the dependence of the amide H-1 and N-15 chemical shifts of CBDN1 upon added sugar, increase from 180 +/- 60 M(-1) for cellotriose to 4 200 +/- 720 M(-1) for cellotetraose, 34 000 +/- 7 600 M(-1) for cellopentaose, and an estimate of 50 000 M(-1) for cellohexaose. This implies that the CBDN1 cellulose-binding site spans approximately five glucosyl units, On the basis of the observed patterns of amide chemical shift changes, the cellooligosaccharides bind along a five-stranded beta-sheet that forms a concave face of the jelly-roll beta-sandwich structure of CBDN1. This beta-sheet contains a strip of hydrophobic side chains flanked on both sides by polar residues, NMR and difference ultraviolet absorbance measurements also demonstrate that tyrosine, but not tryptophan, side chains may be involved in oligosaccharide binding. These results lead to a model in which CBDN1 interacts with soluble cellooligosaccharides and, by inference, with single polysaccharide chains in regions of amorphous cellulose, primarily through hydrogen bonding to the equatorial hydroxyl groups of the pyranose rings. Van der against the apolar side chains may augment binding. CBDN1 stands in marked contrast to previously characterized CBDs that absorb to crystalline cellulose via a flat binding surface dominated by exposed aromatic rings.}, keywords = {BACKBONE H-1, CHEMICAL-SHIFTS, ESCHERICHIA-COLI, EXPRESSION, HIGH-LEVEL, NUCLEAR-MAGNETIC-RESONANCE, PROTEIN, REESEI CELLOBIOHYDROLASE-I, SEQUENCE, TRICHODERMA-REESEI, TRYPTOPHAN RESIDUES}, isbn = {0006-2960}, url = {://A1996VR09800005}, author = {Johnson, P. E. and Tomme, P. and Joshi, M. D. and McIntosh, L. P.} } @article {3551, title = {STRUCTURAL FEATURES OF THE EPSILON-SUBUNIT OF THE ESCHERICHIA-COLI ATP SYNTHASE DETERMINED BY NMR-SPECTROSCOPY}, journal = {Nature Structural Biology}, volume = {2}, number = {11}, year = {1995}, note = {ISI Document Delivery No.: TD554Times Cited: 128Cited Reference Count: 43}, month = {Nov}, pages = {961-967}, type = {Article}, abstract = {The tertiary fold of the epsilon subunit of the Escherichia coli F1F0 ATPsynthase (ECF(1)F(0)) has been determined by two- and three-dimensional heteronuclear (C-13, N-15) NMR spectroscopy. The epsilon subunit exhibits a distinct two domain structure, with the N-terminal 84 residues of the protein forming a 10-stranded beta-structure, and with the C-terminal 48 amino acids arranged as two alpha-helices running antiparallel to one another (two helix hairpin). The beta-domain folds as a beta-sandwich with a hydrophobic interior between the two layers of the sandwich. The C-terminal two-helix hairpin folds back to the N-terminal domain and interacts with one side of the beta-domain. The arrangement of the epsilon subunit in the intact F1F0 ATP synthase involves interaction of the two helix hairpin with the F-1 part, and binding of the open side of the beta-sandwich to the c subunits of the membrane-embedded F-0 part.}, keywords = {AMINO-ACID-SEQUENCE, CRYOELECTRON MICROSCOPY, EXPRESSION, F1, H+-ATPASE, NUCLEOTIDE, PROTEINS, PROTON, RESOLUTION, TRANSLOCATING ADENOSINE-TRIPHOSPHATASE}, isbn = {1072-8368}, url = {://A1995TD55400013}, author = {Wilkens, S. and Dahlquist, F. W. and McIntosh, L. P. and Donaldson, L. W. and Capaldi, R. A.} }