title: Acetylproteomic analysis reveals functional implications of lysine acetylation in human spermatozoa (sperm).
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.
authors: From the NPFPC Key Laboratory of Contraceptives and Devices, Shanghai Institute of Planned Parenthood Research, Institutes of Reproduction and Development.
authors: Heguo
authors: H
authors: Yu
authors: From the NPFPC Key Laboratory of Contraceptives and Devices, Shanghai Institute of Planned Parenthood Research, Institutes of Reproduction and Development.
authors: Hua
authors: H
authors: Diao
authors: 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.
authors: Chunmei
authors: C
authors: Wang
authors: School of Life Sciences, and Molecular and Cell Biology Lab, Fudan University, Shanghai 200032, China.
authors: Yan
authors: Y
authors: Lin
authors: Department of General Surgery, Shanghai First People's Hospital, Medical College, Shanghai Jiaotong University, Shanghai 200080, China, and.
authors: Fudong
authors: F
authors: Yu
authors: Department of Urology, Renji Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai 200001, China.
authors: Hui
authors: H
authors: Lu
authors: School of Life Sciences, and Molecular and Cell Biology Lab, Fudan University, Shanghai 200032, China.
authors: Wei
authors: W
authors: Xu
authors: Department of Urology, Renji Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai 200001, China.
authors: Zheng
authors: Z
authors: Li
authors: From the NPFPC Key Laboratory of Contraceptives and Devices, Shanghai Institute of Planned Parenthood Research, Institutes of Reproduction and Development.
authors: Huijuan
authors: H
authors: Shi
authors: School of Life Sciences, and Molecular and Cell Biology Lab, Fudan University, Shanghai 200032, China, zhaosm@fudan.edu.cn.
authors: Shimin
authors: S
authors: Zhao
authors: 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, zhouych@sibs.ac.cn.
authors: Yuchuan
authors: Y
authors: Zhou
authors: 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, ylzhang@sibcb.ac.cn.
authors: Yonglian
authors: Y
authors: Zhang
mesh_terms: D000107:Acetylation / Q000187:drug effects; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D000918:Antibody Specificity / Q000276:immunology; D002392:Catechin / Q000031:analogs & derivatives / Q000494:pharmacology; D016384:Consensus Sequence; D016572:Cyclosporine / Q000494:pharmacology; D006801:Humans; D008239:Lysine / Q000378:metabolism*; D008297:Male; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D051379:Mice; D008969:Molecular Sequence Data; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D060066:Protein Interaction Maps / Q000187:drug effects; D040901:Proteomics / Q000379:methods*; D012984:Software; D013081:Sperm Motility / Q000187:drug effects; D013094:Spermatozoa / Q000187:drug effects / Q000378:metabolism*; D050996:Voltage-Dependent Anion Channel 2 / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D019151:Peptide Library; D010455:Peptides; C494765:VDAC2 protein, human; D050996:Voltage-Dependent Anion Channel 2; D016572:Cyclosporine; D002392:Catechin; C045651:epigallocatechin gallate; D008239:Lysine