Purification and identification of transglutaminase from mouse coagulating gland and its cross-linking activity among seminal vesicle secretion proteins.
A 75-kDa protein secreted from mouse coagulating gland was purified to homogeneity by a series of isolation steps including ion exchange chromatography on a DEAE-Sephacel column and ion exchange high-performance liquid chromatography on a sulfopropyl column. It was identified to be Type IV transglutaminase (TG(4)), based on the establishment of N-terminal sequences by automated Edman degradation together with partial sequences by MS analysis. Its cross-linking activity was tested on the reduced sample of mouse seminal secretion which contained seven major monomer proteins tentatively designated as SVS I-VII. The enzyme was able to cross-link any of SVS I-III but failed to cross-link the other SVS proteins with a M(r) value less than 14 kDa. SVS I and SVS III showed comparable substrate activity, but were much weaker than SVS II during the TG(4) catalysis.
Tseng
Huan-Chin
HC
Institute of Biochemical Sciences, College of Life Science, National Taiwan University, Taipei, Taiwan.
Lin
Han-Jia
HJ
Sudhakar Gandhi
P S
PS
Wang
Chia-Yih
CY
Chen
Yee-Hsiung
YH
D000818:Animals; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D008297:Male; D051379:Mice; D029607:Seminal Vesicle Secretory Proteins / Q000378:metabolism*; D012669:Seminal Vesicles / Q000737:chemistry*; D013379:Substrate Specificity; D011503:Transglutaminases / Q000302:isolation & purification* / Q000378:metabolism*
D029607:Seminal Vesicle Secretory Proteins; C418702:transglutaminase 4; D011503:Transglutaminases