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排序方式: 共有105条查询结果,搜索用时 31 毫秒
71.
S H Snyder 《Nature》1979,279(5708):13-14
72.
Acute effect of a glucocorticoid on normal human sleep 总被引:3,自引:0,他引:3
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Analysis of yeast protein kinases using protein chips 总被引:26,自引:0,他引:26
Zhu H Klemic JF Chang S Bertone P Casamayor A Klemic KG Smith D Gerstein M Reed MA Snyder M 《Nature genetics》2000,26(3):283-289
We have developed a novel protein chip technology that allows the high-throughput analysis of biochemical activities, and used this approach to analyse nearly all of the protein kinases from Saccharomyces cerevisiae. Protein chips are disposable arrays of microwells in silicone elastomer sheets placed on top of microscope slides. The high density and small size of the wells allows for high-throughput batch processing and simultaneous analysis of many individual samples. Only small amounts of protein are required. Of 122 known and predicted yeast protein kinases, 119 were overexpressed and analysed using 17 different substrates and protein chips. We found many novel activities and that a large number of protein kinases are capable of phosphorylating tyrosine. The tyrosine phosphorylating enzymes often share common amino acid residues that lie near the catalytic region. Thus, our study identified a number of novel features of protein kinases and demonstrates that protein chip technology is useful for high-throughput screening of protein biochemical activity. 相似文献
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Phasing of protein-induced DNA bends in a recombination complex 总被引:26,自引:0,他引:26
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Bibb JA Snyder GL Nishi A Yan Z Meijer L Fienberg AA Tsai LH Kwon YT Girault JA Czernik AJ Huganir RL Hemmings HC Nairn AC Greengard P 《Nature》1999,402(6762):669-671
The physiological state of the cell is controlled by signal transduction mechanisms which regulate the balance between protein kinase and protein phosphatase activities. Here we report that a single protein can, depending on which particular amino-acid residue is phosphorylated, function either as a kinase or phosphatase inhibitor. DARPP-32 (dopamine and cyclic AMP-regulated phospho-protein, relative molecular mass 32,000) is converted into an inhibitor of protein phosphatase 1 when it is phosphorylated by protein kinase A (PKA) at threonine 34. We find that DARPP-32 is converted into an inhibitor of PKA when phosphorylated at threonine 75 by cyclin-dependent kinase 5 (Cdk5). Cdk5 phosphorylates DARPP-32 in vitro and in intact brain cells. Phospho-Thr 75 DARPP-32 inhibits PKA in vitro by a competitive mechanism. Decreasing phospho-Thr 75 DARPP-32 in striatal slices, either by a Cdk5-specific inhibitor or by using genetically altered mice, results in increased dopamine-induced phosphorylation of PKA substrates and augmented peak voltage-gated calcium currents. Thus DARPP-32 is a bifunctional signal transduction molecule which, by distinct mechanisms, controls a serine/threonine kinase and a serine/threonine phosphatase. 相似文献
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Large-scale analysis of the yeast genome by transposon tagging and gene disruption 总被引:67,自引:0,他引:67
Ross-Macdonald P Coelho PS Roemer T Agarwal S Kumar A Jansen R Cheung KH Sheehan A Symoniatis D Umansky L Heidtman M Nelson FK Iwasaki H Hager K Gerstein M Miller P Roeder GS Snyder M 《Nature》1999,402(6760):413-418
Economical methods by which gene function may be analysed on a genomic scale are relatively scarce. To fill this need, we have developed a transposon-tagging strategy for the genome-wide analysis of disruption phenotypes, gene expression and protein localization, and have applied this method to the large-scale analysis of gene function in the budding yeast Saccharomyces cerevisiae. Here we present the largest collection of defined yeast mutants ever generated within a single genetic background--a collection of over 11,000 strains, each carrying a transposon inserted within a region of the genome expressed during vegetative growth and/or sporulation. These insertions affect nearly 2,000 annotated genes, representing about one-third of the 6,200 predicted genes in the yeast genome. We have used this collection to determine disruption phenotypes for nearly 8,000 strains using 20 different growth conditions; the resulting data sets were clustered to identify groups of functionally related genes. We have also identified over 300 previously non-annotated open reading frames and analysed by indirect immunofluorescence over 1,300 transposon-tagged proteins. In total, our study encompasses over 260,000 data points, constituting the largest functional analysis of the yeast genome ever undertaken. 相似文献