Characterization of a membrane-bound arginine-specific serine protease from rat intestinal mucosa.
Previously we isolated and characterized a membrane-bound, arginine-specific serine protease from pig intestinal mucosa [J. Biol. Chem. 269, 32985-32991 (1994)]. For further characterization of this type of enzyme, we cloned a cDNA from rat intestinal mucosa encoding the precursor of a similar protease. The partial amino acid sequences determined for the pig enzyme were found to be shared almost completely by the rat enzyme. The serine protease domain of the rat enzyme, heterologously expressed in Escherichia coli, specifically cleaved Arg (or Lys)-X bonds with a marked preference for Arg-Arg or Arg-Lys, similar to the pig enzyme. The mRNA for the rat enzyme was shown to be distributed mainly in intestine, and the enzyme was detected in the duodenal mucosa as a 70 kDa protein. Immunohistochemical analysis of the small intestinal tissue showed that the enzyme is localized mainly on brushborder membranes.
Kishi
K
K
School of Life Science, Tokyo University of Pharmacy and Life Science, Horinouchi, Hachioji, Tokyo 192-0392, Japan.
Yamazaki
K
K
Yasuda
I
I
Yahagi
N
N
Ichinose
M
M
Tsuchiya
Y
Y
Athauda
S B
SB
Inoue
H
H
Takahashi
K
K
D000595:Amino Acid Sequence; D000818:Animals; D001120:Arginine / Q000378:metabolism*; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D004386:Duodenum / Q000201:enzymology; D004791:Enzyme Inhibitors / Q000378:metabolism; D004947:Esophagus / Q000201:enzymology; D006801:Humans; D007413:Intestinal Mucosa / Q000201:enzymology*; D008297:Male; D008565:Membrane Proteins; D051379:Mice; D008969:Molecular Sequence Data; D051381:Rats; D017208:Rats, Wistar; D017385:Sequence Homology; D012697:Serine Endopeptidases / Q000235:genetics* / Q000378:metabolism*; D013045:Species Specificity; D013347:Subcellular Fractions / Q000201:enzymology; D013379:Substrate Specificity; D013552:Swine; D014018:Tissue Distribution; D014357:Trypsin / Q000378:metabolism
D018076:DNA, Complementary; D004791:Enzyme Inhibitors; D008565:Membrane Proteins; D001120:Arginine; D012697:Serine Endopeptidases; C485593:St14 protein, rat; C485595:matriptase; C485596:ST14 protein, human; C118992:St14 protein, mouse; D014357:Trypsin