Purification and Calcium Dependence of Transglutaminases From Sheep Hair Follicles

Biosci Biotechnol Biochem. 1997 Jul;61(7):1086-90. doi: 10.1271/bbb.61.1086.

Abstract

To study the calcium sensitivity of sheep hair follicle transglutaminase, which was reportedly calcium-independent [H. W. Harding and G.E. Rogers, Biochemistry, 11, 2858-2863 (1972)], the enzyme was purified from a homogenate of merino sheep hair follicles and its calcium dependence was examined. As a result of purification, two types of transglutaminases (DEAE-unabsorbed and absorbed transglutaminase, DU-TG and DA-TG, respectively) were obtained. The molecular mass of DU-TG was 77 and 82 kDa by SDS-PAGE and gel filtration, respectively, while that of DA-TG was 40 and 80 kDa. Each enzyme was obviously calcium dependent and contained (a) cysteine residue(s) in the active site, like other known mammalian transglutaminases. Maximum activation of DU-TG and DA-TG was observed at 1 and 0.1 mM CaCl2, respectively.

MeSH terms

  • Animals
  • Calcium / metabolism*
  • Caseins / chemistry
  • Cross-Linking Reagents / chemistry
  • Dipeptides / chemistry
  • Enzyme Inhibitors / pharmacology
  • Hair Follicle / chemistry
  • Hair Follicle / enzymology*
  • Hydrogen-Ion Concentration
  • Insulin / analogs & derivatives
  • Insulin / chemistry
  • Male
  • Molecular Weight
  • Sheep
  • Temperature
  • Transglutaminases / drug effects
  • Transglutaminases / isolation & purification*
  • Transglutaminases / metabolism*

Substances

  • Caseins
  • Cross-Linking Reagents
  • Dipeptides
  • Enzyme Inhibitors
  • Insulin
  • epsilon-(gamma-glutamyl)-lysine
  • Transglutaminases
  • Calcium