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
ISSN0027-8424
卷号113期号:14页码:E2001-E2010
发表状态已发表
DOI10.1073/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
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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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