Characterization of human recombinant transglutaminase 1 purified from baculovirus-infected insect cells.
Transglutaminase 1 (TGase 1) is required for the formation of a cornified envelope in stratified squamous epithelia. Recombinant human TGase 1 expressed in baculovirus-infected cells was purified in a soluble form at the molecular mass of 92 kDa. Recombinant TGase 1 was susceptible to limited proteolysis by both mu- and m-calpains, the calcium-dependent intracellular cysteine proteases. Although the proteolysis did not induce the elevation of the specific enzyme activity of TGase 1, the requirement of calcium ion in the enzymatic reaction was reduced. Furthermore, the effects of GTP, nitric oxide, and sphingosylphosphocholine, known as regulatory factors for tissue-type isozyme (TGase 2), on the enzymatic activity of TGase 1 were investigated.
Hitomi
K
K
Department of Applied Molecular Biosciences, Graduate School of Bioagricultural Sciences, Nagoya University, Chikusa, Japan. hitomi@agr.nagoya-u.ac.jp
Yamagiwa
Y
Y
Ikura
K
K
Yamanishi
K
K
Maki
M
M
D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D002118:Calcium / Q000378:metabolism; D002154:Calpain / Q000378:metabolism; D018076:DNA, Complementary; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D006160:Guanosine Triphosphate / Q000494:pharmacology; D006801:Humans; D006868:Hydrolysis; D009569:Nitric Oxide / Q000494:pharmacology; D010767:Phosphorylcholine / Q000031:analogs & derivatives* / Q000494:pharmacology; D011994:Recombinant Proteins / Q000009:adverse effects / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D013110:Sphingosine / Q000031:analogs & derivatives* / Q000494:pharmacology; D018411:Spodoptera; D011503:Transglutaminases / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism
D018076:DNA, Complementary; D011994:Recombinant Proteins; C005356:sphingosine phosphorylcholine; D010767:Phosphorylcholine; D009569:Nitric Oxide; D006160:Guanosine Triphosphate; D011503:Transglutaminases; C105804:transglutaminase 1; D002154:Calpain; D013110:Sphingosine; D002118:Calcium