Amino Acids

, Volume 44, Issue 1, pp 209–214

Transglutaminase 2 and Factor XIII catalyze distinct substrates in differentiating osteoblastic cell line: utility of highly reactive substrate peptides

  • Kazuya Watanabe
  • Kanako Tsunoda
  • Miho Itoh
  • Mina Fukui
  • Hitoshi Mori
  • Kiyotaka Hitomi
Original Article

DOI: 10.1007/s00726-011-1131-8

Cite this article as:
Watanabe, K., Tsunoda, K., Itoh, M. et al. Amino Acids (2013) 44: 209. doi:10.1007/s00726-011-1131-8

Abstract

Differentiated osteoblastic cell line, MC3T3-E1 expresses transglutaminase 2 (TG2) and Factor XIII (FXIII). In previous studies, we identified isozyme-specific and highly reactive glutamine-donor substrate peptides (pepF11KA and pepT26) for each isozyme. Using these peptides, we compared the reaction products with lysine-donor substrates for each isozyme in differentiating MC3T3-E1 cells. By this analysis, distinct substrates for the activated TG2 and FXIII were detected in cultured cellular extract. Possible substrates that incorporated biotin-labeled peptides were further purified using streptavidin-affinity chromatography. Several isozyme-specific substrates were identified by mass spectrometry analysis of the purified fractions. These analyses also indicate the benefit of the substrate peptides for obtaining distinct substrates in a reaction mixture where two isozymes co-exist.

Keywords

TransglutaminaseOsteoblastTG2Factor XIII

Abbreviations

Asc-P

l-ascorbic acid 2-phosphate

CBB

Coomassie Brilliant Blue

DTT

Dithiothreitol

TG2

Transglutaminase 2

FXIII

Factor XIII

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Kazuya Watanabe
    • 1
  • Kanako Tsunoda
    • 1
  • Miho Itoh
    • 1
  • Mina Fukui
    • 1
  • Hitoshi Mori
    • 2
  • Kiyotaka Hitomi
    • 1
  1. 1.Department of Applied Molecular BiosciencesGraduate School of Bioagricultural Sciences, Nagoya UniversityNagoyaJapan
  2. 2.Department of Bioengineering SciencesGraduate School of Bioagricultural Sciences, Nagoya UniversityNagoyaJapan