Endogenous substrates for epidermal transglutaminase.
Potential in vivo substrates for epidermal transglutaminase have been isolated and partially characterized in human stratum corneum and new born rat epidermis. [14C]Putrescine and dansylcadaverine were incorporated into epidermal proteins in vitro. Two high molecular weight proteins incorporated the labels in both the rat ahd human homogenates. One of the proteins was too large to enter a 4% sodium dodecyl sulfate-polyacrylamide spacer gel; the other was seen at the interface between the spacer gel and a 10% sodium dodecyl sulphate-polyacrylamide running gel. These proteins were present in a buffer extract, sodium dodecyl sulphate-dithiothreitol extract and NaOH extract. The labels were also incorporated into protein in the insoluble pellet remaining after the afore-mentioned extractions. The incorporation of putrescine and dansylcadaverine was time dependent, and was inhibited by known inhibitors of epidermal transglutaminase. The two high molecular weight proteins had similar amino acid composition, characterized by high glycine, glutamic acid, serine and aspartic acid. The amino acid composition was similar to, although not identical with, the amino acid composition of alpha-keratin proteins. Epidermal homogenates incubated in the presence of transglutaminase showed progressive insolubilization of the protein. This cross-linking was inhibited by putrescine. [14C]Glycine, [14C]histidine and [4C]proline were incorporated into epidermal proteins in newborn rats in vivo. The glycine-labelled protein became progressively more insoluble when incubated in vitro in the presence of transglutaminase. In vitro incubation with transglutaminase had no effect on the histidine-and proline-labelled proteins.
D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D002103:Cadaverine / Q000378:metabolism; D002118:Calcium / Q000494:pharmacology; D003619:Dansyl Compounds / Q000378:metabolism; D004229:Dithiothreitol / Q000494:pharmacology; D004817:Epidermis / Q000201:enzymology* / Q000378:metabolism; D006801:Humans; D008970:Molecular Weight; D011700:Putrescine / Q000378:metabolism; D051381:Rats; D012867:Skin / Q000378:metabolism; D005723:gamma-Glutamyltransferase / Q000378:metabolism*
D000596:Amino Acids; D003619:Dansyl Compounds; D005723:gamma-Glutamyltransferase; D002103:Cadaverine; D002118:Calcium; D004229:Dithiothreitol; D011700:Putrescine