Hierarchies of protein cross-linking in the extracellular matrix: involvement of an egg surface transglutaminase in early stages of fertilization envelope assembly.
The involvement of transglutaminase activity in fertilization envelope (FE) formation was investigated using eggs from the sea urchin, Strongylocentrotus purpuratus. Eggs fertilized in the presence of the transglutaminase inhibitors, putrescine and cadaverine, had disorganized and expanded FEs with inhibition of the characteristic I-T transition. The permeability of the FE was increased by these agents, as revealed by the loss of proteins from the perivitelline space and the appearance of ovoperoxidase activity in supernates from putrescine-treated eggs. [3H]putrescine was incorporated into the FE during fertilization in a reaction catalyzed by an egg surface transglutaminase that could also use dimethylcasein as a substrate in vitelline layer-denuded eggs. Egg secretory products alone had no transglutaminase activity. The cell surface transglutaminase activity was transient and maximal within 4 min of activation. The enzyme was Ca2+ dependent and was inhibited by Zn2+. We conclude that sea urchin egg surface transglutaminase catalyzes an early step in a hierarchy of cross-linking events during FE assembly, one that occurs before ovoperoxidase-mediated dityrosine formation (Foerder, C. A., and B. M. Shapiro. 1977. Proc. Natl. Acad. Sci. USA. 74:4214-4218). Thus it provides a graphic example of the physiological function of a cell surface transglutaminase.
Battaglia
D E
DE
Department of Biochemistry, University of Washington, Seattle 98195.
Shapiro
B M
BM
D000818:Animals; D005109:Extracellular Matrix / Q000648:ultrastructure*; D005306:Fertilization; D008855:Microscopy, Electron, Scanning; D010544:Peroxidases / Q000502:physiology; D011700:Putrescine / Q000494:pharmacology; D012617:Sea Urchins / Q000201:enzymology; D011503:Transglutaminases / Q000502:physiology*; D015053:Zygote / Q000648:ultrastructure*
D010544:Peroxidases; D011503:Transglutaminases; D011700:Putrescine