Purification, characterization, and relation to bikunin of rat urinary trypsin inhibitors.
Two forms of urinary trypsin inhibitor (UTI-1 and UTI-2) were purified from pooled urine of normal male rats to apparent homogeneity by salting out, affinity chromatography, gel filtration, and reverse-phase HPLC. UTIs-1 and 2 were shown to be thermostable glycoproteins with the respective molecular weights of 22,000 and 18,000 estimated by SDS-PAGE. These inhibitors combined with bovine trypsin in a 1:1 molar ratio: the Kd values were 2.5 x 10(-10) and 2.3 x 10(-10) M, respectively. Amino acid composition and sequence analysis indicated that UTI-1 corresponded to rat bikunin of which the amino acid sequence was deduced from a rat liver cDNA clone encoding alpha1-microglobulin [Lindqvist et al. (1992), Biochim. Biophys. Acta 1130, 63-67] except that the protein sequence seemed to lack C-terminal serine, and UTI-2 corresponded to UTI-1 lacking N-terminal 21 amino acid residues.
Kurata
A
A
College of Human Ecology, Kyoto Koka Women's University, Japan.
Ohi
K
K
Sato
K
K
Tashiro
M
M
D000595:Amino Acid Sequence; D000818:Animals; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D006023:Glycoproteins / Q000737:chemistry / Q000302:isolation & purification*; D008297:Male; D008562:Membrane Glycoproteins / Q000737:chemistry*; D008969:Molecular Sequence Data; D020033:Protein Isoforms / Q000737:chemistry / Q000302:isolation & purification*; D051381:Rats; D017208:Rats, Wistar; D017386:Sequence Homology, Amino Acid; D014360:Trypsin Inhibitor, Kunitz Soybean
D006023:Glycoproteins; D008562:Membrane Glycoproteins; D020033:Protein Isoforms; C109148:SPINT2 protein, human; D014360:Trypsin Inhibitor, Kunitz Soybean; C028665:urinastatin