Site-specific cross-linking of functional proteins by transglutamination

Noriho Kamiya, Takeshi Takazawa, Tsutomu Tanaka, Hiroshi Ueda, Teruyuki Nagamune

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

Microbial transglutaminase (MTG), an enzyme that works in posttranslational modification of proteins, has been applied to in vitro preparation of a bi-functional fusion protein from separately obtained recombinant proteins. A quite simple strategy in which a specific peptidyl linker recognizable by MTG is fused to the N-terminus of proteins of interest has been verified in the preparation of a bi-functional fusion protein using an antibody variable domain (single chain Fv of anti-hen-egg white lysozyme antibody, scFv) and a fluorescent protein (enhanced yellow fluorescent protein, EYFP). The resultant peptidyl linker-fused proteins were readily cross-linked by MTG to only give the heterodimer (i.e. scFv-EYFP fusion protein), suggesting that the reaction proceeded in highly specific manner. As the fusion protein exhibited sufficient bi-functionality in fluorescence immunoassay (FIA), this work shows for the first time a successful enzymatic preparation of a bi-functional fusion protein through recombinantly incorporated specific peptidyl linkers into target proteins.

Original languageEnglish
Pages (from-to)492-496
Number of pages5
JournalEnzyme and Microbial Technology
Volume33
Issue number4
DOIs
Publication statusPublished - Sep 10 2003

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Proteins
Fusion reactions
Transglutaminases
Single-Chain Antibodies
Antibodies
Enzymes
Recombinant proteins
Egg White
Post Translational Protein Processing
Muramidase
Immunoassay
Recombinant Proteins
Fluorescence

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Biochemistry
  • Applied Microbiology and Biotechnology

Cite this

Site-specific cross-linking of functional proteins by transglutamination. / Kamiya, Noriho; Takazawa, Takeshi; Tanaka, Tsutomu; Ueda, Hiroshi; Nagamune, Teruyuki.

In: Enzyme and Microbial Technology, Vol. 33, No. 4, 10.09.2003, p. 492-496.

Research output: Contribution to journalArticle

Kamiya, Noriho ; Takazawa, Takeshi ; Tanaka, Tsutomu ; Ueda, Hiroshi ; Nagamune, Teruyuki. / Site-specific cross-linking of functional proteins by transglutamination. In: Enzyme and Microbial Technology. 2003 ; Vol. 33, No. 4. pp. 492-496.
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