title: Testis-specific serine/threonine protein kinase 4 (Tssk4) phosphorylates Odf2 at Ser-76.
abstract: As a member of the testis-specific serine/threonine protein kinase (TSSK) family, Tssk4 is exclusively expressed in the testis and plays an essential role in male fertility. We previously reported that Tssk4 can associate with and phosphorylate Odf2, but the phosphorylation site is still unknown. Here we confirm that the C-terminal region (amino acids 214-638) of Odf2 is required for association with Tssk4. Furthermore, to identify the site at which Tssk4 phosphorylates Odf2, we generated several Odf2 point mutants (Ser/Thr/Lys to Ala) and identified serine 76 of Odf2 as one of the phosphorylation sites. In vivo, phosphorylated Odf2 was evaluated in mouse sperm using a specific phospho-Ser-76 Odf2 antibody and LC-MS/MS. These findings are the first to demonstrate the phosphorylation site in Odf2 by Tssk4, providing essential clues regarding the function of Tssk4 in regulating sperm motility and/or structure and thus male fertility.
authors.affiliation: Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education, School of Pharmaceutical Sciences, and Medical Research Institute, Wuhan University, Wuhan, 430072, P.R.China.
authors.forename: Xiaoli
authors.initials: X
authors.lastname: Wang
authors.affiliation: Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education, School of Pharmaceutical Sciences, and Medical Research Institute, Wuhan University, Wuhan, 430072, P.R.China.|Taihe Hospital, Hubei University of Medicine, Shiyan, Hubei, China.
authors.forename: Han
authors.initials: H
authors.lastname: Li
authors.affiliation: State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Fudan University, 220 Handan Road, Shanghai 200433, P.R. China.
authors.forename: Guolong
authors.initials: G
authors.lastname: Fu
authors.affiliation: Taihe Hospital, Hubei University of Medicine, Shiyan, Hubei, China.
authors.forename: Yunfu
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Taihe Hospital, Hubei University of Medicine, Shiyan, Hubei, China.
authors.forename: Shiming
authors.initials: S
authors.lastname: Du
authors.affiliation: State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Fudan University, 220 Handan Road, Shanghai 200433, P.R. China.
authors.forename: Long
authors.initials: L
authors.lastname: Yu
authors.affiliation: State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Fudan University, 220 Handan Road, Shanghai 200433, P.R. China.
authors.forename: Youheng
authors.initials: Y
authors.lastname: Wei
authors.affiliation: Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education, School of Pharmaceutical Sciences, and Medical Research Institute, Wuhan University, Wuhan, 430072, P.R.China.
authors.forename: Shi
authors.initials: S
authors.lastname: Chen
mesh_terms: D000818:Animals; D006360:Heat-Shock Proteins / Q000378:metabolism*; D008297:Male; D008810:Mice, Inbred C57BL; D010766:Phosphorylation; D017354:Point Mutation; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism*; D012694:Serine / Q000378:metabolism*; D013081:Sperm Motility; D013094:Spermatozoa / Q000378:metabolism
chemical_list: D006360:Heat-Shock Proteins; C490106:Odf2 protein, mouse; D012694:Serine; D017346:Protein Serine-Threonine Kinases