TY - JOUR T1 - Substrate profiling of the metalloproteinase ovastacin – Implications for its physiological function in mammalian fertilization JF - bioRxiv DO - 10.1101/2022.12.06.519252 SP - 2022.12.06.519252 AU - Matthias Felten AU - Ute Distler AU - Nele v. Wiegen AU - Mateusz Łącki AU - Christian Behl AU - Stefan Tenzer AU - Walter Stöcker AU - Hagen Körschgen Y1 - 2022/01/01 UR - http://biorxiv.org/content/early/2022/12/06/2022.12.06.519252.abstract N2 - The metalloproteinase ovastacin is released by the mammalian egg upon fertilization and cleaves a distinct peptide bond in zona pellucida protein 2, a component of the enveloping extracellular matrix. This limited proteolysis causes zona pellucida hardening, abolishes sperm binding and thereby regulates fertility. Accordingly, this process is tightly controlled by the plasma protein fetuin-B, an endogenous competitive inhibitor. At present, little is known about how the cleavage characteristics of ovastacin differ from closely related proteases. Physiological implications of ovastacin beyond ZP2 cleavage are still obscure. In this study, we employed N-terminal amine isotopic labeling of substrates (N-TAILS) contained in the secretome of mouse embryonic fibroblasts to elucidate the substrate specificity and the precise cleavage site specificity. Furthermore, we were able to unravel the physicochemical properties governing enzyme-substrate interactions. Eventually, we identified several potential physiological substrates with significance for mammalian fertilization. These data suggest that ovastacin might regulate sperm-oocyte interaction and fertility beyond zona pellucida hardening.Competing Interest StatementThe authors have declared no competing interest. ER -