Purification and calcium dependence of transglutaminases from sheep hair follicles.
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.
Kumazawa
Y
Y
Ajinomoto Co., Inc., Food Development & Research Laboratories, Kanagawa, Japan.
Ohtsuka
T
T
Ninomiya
D
D
Seguro
K
K
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002364:Caseins / Q000737:chemistry; D003432:Cross-Linking Reagents / Q000737:chemistry; D004151:Dipeptides / Q000737:chemistry; D004791:Enzyme Inhibitors / Q000494:pharmacology; D018859:Hair Follicle / Q000737:chemistry / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D007328:Insulin / Q000031:analogs & derivatives / Q000737:chemistry; D008297:Male; D008970:Molecular Weight; D012756:Sheep; D013696:Temperature; D011503:Transglutaminases / Q000187:drug effects / Q000302:isolation & purification* / Q000378:metabolism*
D002364:Caseins; D003432:Cross-Linking Reagents; D004151:Dipeptides; D004791:Enzyme Inhibitors; D007328:Insulin; C001299:epsilon-(gamma-glutamyl)-lysine; D011503:Transglutaminases; D002118:Calcium