Expression of human transglutaminase C (TGase II) in yeast saccharomyces cereisiae

An effect of a domain from carboxyl terminal deletion of the enzyme

Sang K. Woo, Sun M. Jung, Sang K. Rhee, Byung-Yoon Ahn, Hee Chul Kim

Research output: Contribution to journalArticle

Abstract

In an effort to understand the role of the conserved domain and of the heterologous one-third part of the carboxy terminal domain of transglutaminase C (TGase II), attempts were made to express TGase II cDNA of human origin in yeast Saccharomyces cerevisiae as in a full-length form as well as in a form of C-terminal truncation. The 2μ-based expression plasmids which contained the TGase II cDNA under the gal inducible promoter were introduced into yeast and the maintenance of the full-length and truncated form of the TGase II gene plasmids were confirmed by Southern blot. The expression of the TGase II gene was analysed by reverse transcription polymerase chain reaction (RT-PCR), and western blot analyses. As assayed by [1,414C]-putrescine incorporation into succinylated casein, the full-lenth as well as the truncated forms of recombinant TGase II showed some catalytic activity. These results indicate that the N-terminal homologous domain of human TGase II retains a catalytically active domain. The level of TGase II expressed in yeast, however, was far lower than satisfactory and other expression system should be sought further chracterization of the enzyme. The negative effect of TGase II on the growth of yeast is interesting with respect to the physiological effect of TGase II in cornification of epidermal keratinocytes.

Original languageEnglish
Pages (from-to)290-298
Number of pages9
JournalKorean Journal of Applied Microbiology and Biotechnology
Volume24
Issue number3
Publication statusPublished - 1996 Dec 1

Fingerprint

Saccharomyces
Yeasts
Enzymes
Plasmids
transglutaminase 2
Complementary DNA
Putrescine
Southern Blotting
Caseins
Keratinocytes
Genes
Reverse Transcription
Saccharomyces cerevisiae
Western Blotting
Maintenance

Keywords

  • Cross-linking
  • Transglutaminase C (TGase II)

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology

Cite this

Expression of human transglutaminase C (TGase II) in yeast saccharomyces cereisiae : An effect of a domain from carboxyl terminal deletion of the enzyme. / Woo, Sang K.; Jung, Sun M.; Rhee, Sang K.; Ahn, Byung-Yoon; Kim, Hee Chul.

In: Korean Journal of Applied Microbiology and Biotechnology, Vol. 24, No. 3, 01.12.1996, p. 290-298.

Research output: Contribution to journalArticle

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