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Quantitative MS-based enzymology of caspases reveals distinct protein substrate specificities, hierarchies, and cellular roles | |
Julien, Olivier1; Zhuang, Min1,3; Wiita, Arun P.1,4; O'Donoghue, Anthony J.1,5; Knudsen, Giselle M.1; Craik, Charles S.1; Wells, James A.1,2 | |
2016-04-05 | |
发表期刊 | PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA |
ISSN | 0027-8424 |
卷号 | 113期号:14页码:E2001-E2010 |
发表状态 | 已发表 |
DOI | 10.1073/pnas.1524900113 |
摘要 | Proteases constitute the largest enzyme family, yet their biological roles are obscured by our rudimentary understanding of their cellular substrates. There are 12 human caspases that play crucial roles in inflammation and cell differentiation and drive the terminal stages of cell death. Recent N-terminomics technologies have begun to enumerate the diverse substrates individual caspases can cleave in complex cell lysates. It is clear that many caspases have shared substrates; however, few data exist about the catalytic efficiencies (kcat/KM) of these substrates, which is critical to understanding their true substrate preferences. In this study, we use quantitative MS to determine the catalytic efficiencies for hundreds of natural protease substrates in cellular lysate for two understudied members: caspase-2 and caspase-6. Most substrates are new, and the cleavage rates vary up to 500-fold. We compare the cleavage rates for common substrateswith those found for caspase-3, caspase-7, and caspase-8, involved in apoptosis. There is little correlation in catalytic efficiencies among the five caspases, suggesting each has a unique set of preferred substrates, and thus more specialized roles than previously understood. We synthesized peptide substrates on the basis of protein cleavage sites and found similar catalytic efficiencies between the protein and peptide substrates. These data suggest the rates of proteolysis are dominated more by local primary sequence, and less by the tertiary protein fold. Our studies highlight that global quantitative rate analysis for posttranslational modification enzymes in complex milieus for native substrates is critical to better define their functions and relative sequence of events. |
关键词 | N-terminomics caspase proteolysis proteomics apoptosis |
收录类别 | SCI |
语种 | 英语 |
资助项目 | National Institutes of Health[R21CA186007] ; National Institutes of Health[R01GM081051] ; National Institutes of Health[R01GM097316] ; National Institutes of Health[R01CA154802] |
WOS研究方向 | Science & Technology - Other Topics |
WOS类目 | Multidisciplinary Sciences |
WOS记录号 | WOS:000373354000010 |
出版者 | NATL ACAD SCIENCES |
WOS关键词 | COMBINATORIAL APPROACH ; GLOBAL IDENTIFICATION ; TARGETED PROTEOMICS ; PROTEOLYTIC EVENTS ; CLEAVAGE ; ACTIVATION ; APOPTOSIS ; PROTEASES ; TOPOGRAPHY ; MECHANISMS |
原始文献类型 | Article |
引用统计 | |
文献类型 | 期刊论文 |
条目标识符 | https://kms.shanghaitech.edu.cn/handle/2MSLDSTB/1875 |
专题 | 生命科学与技术学院_PI研究组_庄敏组 |
通讯作者 | Wells, James A. |
作者单位 | 1.Univ Calif San Francisco, Dept Pharmaceut Chem, San Francisco, CA 94143 USA 2.Univ Calif San Francisco, Dept Cellular & Mol Pharmacol, San Francisco, CA 94143 USA 3.ShanghaiTech Univ, Sch Life Sci & Technol, Shanghai 201210, Peoples R China 4.Univ Calif San Francisco, Dept Lab Med, San Francisco, CA 94143 USA 5.Univ Calif San Diego, Skaggs Sch Pharm & Pharmaceut Sci, San Diego, CA 92093 USA |
推荐引用方式 GB/T 7714 | Julien, Olivier,Zhuang, Min,Wiita, Arun P.,et al. Quantitative MS-based enzymology of caspases reveals distinct protein substrate specificities, hierarchies, and cellular roles[J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA,2016,113(14):E2001-E2010. |
APA | Julien, Olivier.,Zhuang, Min.,Wiita, Arun P..,O'Donoghue, Anthony J..,Knudsen, Giselle M..,...&Wells, James A..(2016).Quantitative MS-based enzymology of caspases reveals distinct protein substrate specificities, hierarchies, and cellular roles.PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA,113(14),E2001-E2010. |
MLA | Julien, Olivier,et al."Quantitative MS-based enzymology of caspases reveals distinct protein substrate specificities, hierarchies, and cellular roles".PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 113.14(2016):E2001-E2010. |
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