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PMID: 25680958 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Acetylproteomic analysis reveals functional implications of lysine acetylation in human spermatozoa (sperm).

Molecular & cellular proteomics : MCP ·Vol. 14 ·No. 4 ·2015-04-00 ·Pages 1009-23

Yu H, Diao H, Wang C, Lin Y, Yu F, Lu H, Xu W, Li Z, Shi H, Zhao S, Zhou Y, Zhang Y

Abstract

Male infertility is a medical condition that has been on the rise globally. Lysine acetylation of human sperm, an essential posttranslational modification involved in the etiology of sperm abnormality, is not fully understood. Therefore, we first generated a qualified pan-anti-acetyllysine monoclonal antibody to characterize the global lysine acetylation of uncapacitated normal human sperm with a proteomics approach. With high enrichment ratios that were up to 31%, 973 lysine-acetylated sites that matched to 456 human sperm proteins, including 671 novel lysine acetylation sites and 205 novel lysine-acetylated proteins, were identified. These proteins exhibited conserved motifs XXXKYXXX, XXXKFXXX, and XXXKHXXX, were annotated to function in multiple metabolic processes, and were localized predominantly in the mitochondrion and cytoplasmic fractions. Between the uncapacitated and capacitated sperm, different acetylation profiles in regard to functional proteins involved in sperm capacitation, sperm-egg recognition, sperm-egg plasma fusion, and fertilization were observed, indicating that acetylation of functional proteins may be required during sperm capacitation. Bioinformatics analysis revealed association of acetylated proteins with diseases and drugs. Novel acetylation of voltage-dependent anion channel proteins was also found. With clinical sperm samples, we observed differed lysine acetyltransferases and lysine deacetylases expression between normal sperm and abnormal sperm of asthenospermia or necrospermia. Furthermore, with sperm samples impaired by epigallocatechin gallate to mimic asthenospermia, we observed that inhibition of sperm motility was partly through the blockade of voltage-dependent anion channel 2 Lys-74 acetylation combined with reduced ATP levels and mitochondrial membrane potential. Taken together, we obtained a qualified pan-anti-acetyllysine monoclonal antibody, analyzed the acetylproteome of uncapacitated human sperm, and revealed associations between functional protein acetylation and sperm functions.

MeSH Terms
Acetylation/drug effects Amino Acid Motifs Amino Acid Sequence Animals Antibodies, Monoclonal/metabolism Antibody Specificity/immunology Catechin/analogs & derivatives,pharmacology Consensus Sequence Cyclosporine/pharmacology Humans Lysine/metabolism Male Membrane Potential, Mitochondrial/drug effects Mice Molecular Sequence Data Peptide Library Peptides/chemistry,metabolism Protein Interaction Maps/drug effects Proteomics/methods Software Sperm Motility/drug effects Spermatozoa/drug effects,metabolism Voltage-Dependent Anion Channel 2/metabolism
Chemicals
Antibodies, Monoclonal Peptide Library Peptides VDAC2 protein, human Voltage-Dependent Anion Channel 2 Cyclosporine Catechin epigallocatechin gallate Lysine
Authors & Affiliations
12 authors, click to expand affiliations / ORCID
Yu Heguo
From the NPFPC Key Laboratory of Contraceptives and Devices, Shanghai Institute of Planned Parenthood Research, Institutes of Reproduction and Development.
Diao Hua
From the NPFPC Key Laboratory of Contraceptives and Devices, Shanghai Institute of Planned Parenthood Research, Institutes of Reproduction and Development.
Wang Chunmei
Shanghai Key Laboratory for Molecular Andrology, State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
Lin Yan
School of Life Sciences, and Molecular and Cell Biology Lab, Fudan University, Shanghai 200032, China.
Yu Fudong
Department of General Surgery, Shanghai First People's Hospital, Medical College, Shanghai Jiaotong University, Shanghai 200080, China, and.
Lu Hui
Department of Urology, Renji Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai 200001, China.
Xu Wei
School of Life Sciences, and Molecular and Cell Biology Lab, Fudan University, Shanghai 200032, China.
Li Zheng
Department of Urology, Renji Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai 200001, China.
Shi Huijuan
From the NPFPC Key Laboratory of Contraceptives and Devices, Shanghai Institute of Planned Parenthood Research, Institutes of Reproduction and Development.
Zhao Shimin
School of Life Sciences, and Molecular and Cell Biology Lab, Fudan University, Shanghai 200032, China, [email protected].
Zhou Yuchuan
Shanghai Key Laboratory for Molecular Andrology, State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China, [email protected].
Zhang Yonglian
From the NPFPC Key Laboratory of Contraceptives and Devices, Shanghai Institute of Planned Parenthood Research, Institutes of Reproduction and Development, Shanghai Key Laboratory for Molecular Andrology, State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China, [email protected].
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Article Info
Journal
Molecular & cellular proteomics : MCP
Abbr.
Mol Cell Proteomics
ISSN
1535-9484
DOI
10.1074/mcp.M114.041384
Email
[email protected]
[email protected]
[email protected]
Published
2015-04-00
Epub
2015-00-13
Pages
1009-23
Language
English
Region
United States
NLM ID
101125647
PMCID
PMC4390248
Subset
IM
External Links
PubMed source DOI: 10.1074/mcp.M114.041384 PMC Full Text View journal details
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