P-085 Reactive molecules drive sperm capacitation via protein posttranslational modifications

Abstract Study question Are S-sulfenylation and S-nitrosylation, two essential posttranslational modifications (PTMs), increased during sperm capacitation? Which proteins are subjected to S-sulfenylation? Summary answer S-nitrosylation increase capacitation. affects certain proteins, including A-kinase anchor protein 13, Fibrilline-1, Spermatogenesis-associated 7. What is known already There a significant in reactive oxygen species (ROS) nitrogen (RNS) production within In contrast tyrosine phosphorylation, well-established mark of capacitation, ROS, RNS lead PTMs themselves modify thiol groups cysteines. Despite these facts, the dynamics ROS/RNS-derived capacitation remain unknown. Therefore, we consider both as novel associated with design, size, duration C57BL6 mouse were isolated from caudae epididymae 12-20-week-old males (n = 5). The study compared non-capacitated capacitated spermatozoa using proteomic profiling. Participants/materials, setting, methods Sperm extracted into RIPA buffer. lysate was incubated 5mM dimedone, hapten that specifically labels S-sulfenylated immunogenically detected an anti-dimedone antibody (Merk, Germany). Alternatively, S-nitrosylated by anti-nitrosocysteine visualized western blotting. simultaneously identified mass spectrometry. Main results role chance We demonstrated along addition, positively correlated phosphorylation. Using spectrometry, mostly belonging cytoskeleton: 7, Rab-like 2A. Limitations, reasons for caution limitation this instability PTMs. hypothetically arise cease sample preparation. experiment limited one animal model. Further experiments should be performed on other mammalian models. Wider implications findings Studying their mechanism action may improve diagnosis male infertility. Further, it could bring approach modulate fertilizing ability used vitro fertilization. Trial registration number 260 536