Discovery of a human testis-specific protein complex TEX101-DPEP3 and selection of its disrupting antibodies
Discovery of a human testis-specific protein complex TEX101-DPEP3 and selection of its disrupting antibodies
SUMMARY TEX101 is a testis-specific protein expressed exclusively in male germ cells and is a validated biomarker of male infertility. Studies in mice suggest that TEX101 is a cell-surface chaperone which regulates, through protein-protein interactions, the maturation of proteins involved in spermatozoa transit and oocyte binding. Male TEX101-null mice are sterile. Here, we identified by co-immunoprecipitation-mass spectrometry the interactome of human TEX101 in testicular tissues and spermatozoa. The testis-specific cell-surface dipeptidase 3 (DPEP3) emerged as the top hit. We further validated the TEX101-DPEP3 complex by using hybrid immunoassays. Combinations of antibodies recognizing different epitopes of TEX101 and DPEP3 facilitated development of a simple immunoassay to screen for disruptors of TEX101-DPEP3 complex. As a proof-of-a-concept, we demonstrated that anti-TEX101 antibody T4 disrupted the native TEX101-DPEP3 complex. Disrupting antibodies may be used to study the human TEX101-DPEP3 complex, and to develop modulators for male fertility. Non-standard abbreviationsTEX101Testis-expressed sequence 101 proteinDPEP3Dipeptidase 3AC-MSAffinity capture-mass spectrometryco-IP-MSCoimmunoprecipitation-mass spectrometryGPIGlycosylphosphatidylinositolLFQLabel-free quantificationmAbMonoclonal antibodyNHSN-hydroxysuccinimidePRMParallel reaction monitoringPTMPosttranslational modificationSPSeminal plasmaSRMSelected reaction monitoringFDRfalse detection rate
Schiza Christina、Panteleli Efstratia、Jarvi Keith、Drabovich Andrei P.、Diamandis Eleftherios P.、Korbakis Dimitrios
Department of Laboratory Medicine and Pathobiology, University of Toronto||Department of Pathology and Laboratory Medicine, Mount Sinai HospitalDepartment of Clinical Biochemistry, University Health NetworkLunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital||Department of Surgery, Division of Urology, Mount Sinai HospitalDepartment of Laboratory Medicine and Pathobiology, University of Toronto||Department of Pathology and Laboratory Medicine, Mount Sinai Hospital||Department of Clinical Biochemistry, University Health NetworkDepartment of Laboratory Medicine and Pathobiology, University of Toronto||Department of Pathology and Laboratory Medicine, Mount Sinai Hospital||Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital||Department of Clinical Biochemistry, University Health NetworkDepartment of Laboratory Medicine and Pathobiology, University of Toronto||Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital
基础医学生物科学研究方法、生物科学研究技术生理学
TEX101testis-expressed sequence 101 proteinDPEP3dipeptidase 3affinity purification-mass spectrometryinteractomeprotein-protein interactionstestis-specific proteinsspermatozoadisrupting antibodies
Schiza Christina,Panteleli Efstratia,Jarvi Keith,Drabovich Andrei P.,Diamandis Eleftherios P.,Korbakis Dimitrios.Discovery of a human testis-specific protein complex TEX101-DPEP3 and selection of its disrupting antibodies[EB/OL].(2025-03-28)[2025-05-13].https://www.biorxiv.org/content/10.1101/315713.点此复制
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