2003 Fiscal Year Final Research Report Summary

Mechanism and role of croslinking to cell membrane of cystatin A

Research Project

Project/Area Number	13670860
Research Category	Grant-in-Aid for Scientific Research (C)
Allocation Type	Single-year Grants
Section	一般
Research Field	Dermatology
Research Institution	Asahikawa Medical College
Principal Investigator	TAKAHASHI Hidetoshi Asahikawa Medical College, Assistant professor, 医学部, 講師 (00216748)
Project Period (FY)	2001 – 2003
Keywords	cystatin A / cornified envelope / 44Lys / cross-linking / keratinocyte / substitution / mutation / differentiation
Research Abstract	We produced wild and mutant cystatin A gene, which were substituted the amino acids by in vitro mutagenesis kit. These products were cross-linked each other by the transglutaminase, then were electrohoresed by SDS-page. Only one product, which was substituted Lys at 44 amino acid to Thr, could not cross-link. Next these genes were introduced into the human keratinocytes. Then the keratinoctes were induced to form cornified cell envelope by calcium ionophore. We could observe the incorporation of cystatin A into inner surface of keratinocytes, which was induced Lys44Thr gene. The present study demonstrated that Lys at 44 amino acid is important for cross-linkinking to cell membrane.

(2 results)