耶魯大學(xué)科學(xué)家Michael Snyder等首次通過研究酵母中的蛋白磷酸化網(wǎng)絡(luò)，描繪出控制高等生物的細(xì)胞功能的蛋白激酶的信號網(wǎng)絡(luò),。研究人員指出本研究是一項重大的突破，解釋了蛋白激酶這一調(diào)控因子如何使蛋白質(zhì)在不同類型的細(xì)胞中發(fā)揮作用,。盡管目前蛋白激酶已經(jīng)作為一種重要抗癌藥物的靶位點,，然而到目前為止受激酶調(diào)控的蛋白質(zhì)還很難確定。
由細(xì)胞分子生物學(xué)家Michael Snyder負(fù)責(zé)的研究小組觀察了酵母細(xì)胞不同蛋白質(zhì)的表達(dá),、相互作用以及細(xì)胞中的活性蛋白成分,。作為調(diào)控因子的蛋白激酶通過對靶蛋白的磷酸化從而使其發(fā)揮作用。這一過程最終會導(dǎo)致蛋白質(zhì)的活性發(fā)生改變,。據(jù)估計,，約30％的蛋白質(zhì)都通過這一途徑進(jìn)行調(diào)控。
生物谷認(rèn)為,，采用酵母作為大規(guī)模蛋白質(zhì)間相互作用,，或激酶網(wǎng)絡(luò)模型是近年來研究趨勢，先前有報道：酵母膜表面蛋白相互作用的大規(guī)模研究,、利用RNAi技術(shù)對生命活動中的激酶進(jìn)行大規(guī)模研究以及酵母的大規(guī)模功能蛋白學(xué)研究結(jié)果等,，這些大規(guī)模研究有助于了解生命現(xiàn)象的復(fù)雜過程，也有助于理解生命的活動的本質(zhì)特征,。
這項研究結(jié)果發(fā)表在最近一期的《自然》雜志上,。
a, Overall scheme to identify kinase substrates. Each kinase was overexpressed, purified and assayed on protein chips containing about 4,400 proteins spotted in duplicate. b, Kinase assays on protein chips. Two protein chips were used for every kinase assayed. In addition, two protein chips were probed in the absence of kinase to identify proteins on the chip that autophosphorylate. Commercial kinases were spotted at many defined locations, shown in the four corners of the two boxes on the right; these kinases autophosphorylated in our assay and served as landmarks for the identification of phosphorylation signals. The slide on the left is a representative slide probed with anti-GST antibodies indicating the amount of fusion protein present on the proteome slide. c, In vivo validation of targets identified on the proteome microarray: ARK1 (i), SWE1 (ii), HSL1 (iii), BCK1 (iv), STE20 (v) and PRK1 (vi) were deleted from the TAP-tagged strains indicated. From the kinase-deleted strains, the tagged proteins were purified and their phosphorylation status compared with wild-type tagged proteins. Immunoblots were probed with anti-phosphothreonine antibody (i, top panel), anti-phosphotyrosine antibody (ii, top panel) or anti-phosphoserine antibody (iv, top panel). In addition, protein isoforms and protein levels were monitored with anti-CBP antibody (i–vi).
a, A map showing the connections between kinases and substrates. In all, 87 different kinases (red dots) and 1,325 substrates (blue dots) are represented in the map. b, Global localization data can be used to identify only those phosphorylation events occurring between proteins of the same cellular compartment. c, Functional data can be used to identify substrates with similar functions to those of the kinases phosphorylating them.
相關(guān)研究報道
·《Nature》：大規(guī)模繪制基因圖譜的新方法
·《任小二快報》：人類基因組的大規(guī)模拷貝數(shù)的多態(tài)性
·大規(guī)模RNAi技術(shù)首次在哺乳動物中應(yīng)用
·中心粒粘附的相關(guān)基因的大規(guī)模篩選
·用RNAi技術(shù)大規(guī)模研究基因功能
·酵母的大規(guī)模功能蛋白學(xué)研究結(jié)果
·酵母膜表面蛋白相互作用的大規(guī)模研究
·利用RNAi技術(shù)對生命活動中的激酶進(jìn)行大規(guī)模研究
·酵母的大規(guī)模功能蛋白學(xué)研究結(jié)果
原始出處：
Jason Ptacek, Geeta Devgan, Gregory Michaud, Heng Zhu, Xiaowei Zhu, Joseph Fasolo, Hong Guo, Ghil Jona, Ashton Breitkreutz, Richelle Sopko, Rhonda R. McCartney, Martin C. Schmidt, Najma Rachidi, Soo-Jung Lee, Angie S. Mah, Lihao Meng, Michael J. R. Stark, David F. Stern, Claudio De Virgilio, Mike Tyers, Brenda Andrews, Mark Gerstein, Barry Schweitzer, Paul F. Predki and Michael Snyder. Global analysis of protein phosphorylation in yeast. Nature 438, 679-684 (1 December 2005)
實驗室主頁：
Michael Snyder, Ph.D. [email protected]
Professor and Chairman of Molecular, Cellular, and Developmental Biology;
Professor of Molecular Biophysics & Biochemistry
Contact and Mailing Information
B.A. Chemistry and Biology;
University of Rochester, N.Y. 1977;
Ph.D. Biology California Institute
of Technology 1983;
Helen Hay Whitney Fellow;
Stanford University 1982-86;
Joined Yale Faculty 1986;
United Scleroderma Foundation Award;
Pew Scholar Award 1987-91.
Research Interests:
Molecular Genetics/Cell Biology
