Cloning and characterization of mouse klk27, a novel tissue kallikrein expressed in testicular Leydig cells and exhibiting chymotrypsin-like specificity.
A cDNA clone of a new mouse tissue kallikrein, designated mKlk27, was isolated from an adult mouse testis cDNA library. mKlk27 was expressed in the submaxillary glands and testis of the mouse. In testis, mKlk27 gene was expressed exclusively in the Leydig cells of the adult mouse. Active recombinant mKlk27 exhibited chymotrypsin-like cleavage specificity. A single amino-acid substitution of Gly for Asp at position 209 in mKlk27 resulted in complete loss of its chymotryptic activity but acquisition of tryptic activity. mKlk27 effectively hydrolyzed casein, gelatin and fibronectin. Insulin-like growth factor binding protein-3 was also hydrolyzed by recombinant mKlk27. These results suggest that mKlk27 plays an important role in association with the function of the adult mouse testis.
Matsui
H
H
Division of Biological Sciences, Hokkaido University, Sapporo, Japan.
Moriyama
A
A
Takahashi
T
T
D000595:Amino Acid Sequence; D000818:Animals; D001224:Aspartic Acid / Q000737:chemistry; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D002364:Caseins / Q000378:metabolism; D002918:Chymotrypsin / Q000378:metabolism*; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005353:Fibronectins / Q000378:metabolism; D005780:Gelatin / Q000378:metabolism; D015723:Gene Library; D005998:Glycine / Q000737:chemistry; D006868:Hydrolysis; D017403:In Situ Hybridization; D018972:Insulin-Like Growth Factor Binding Protein 3 / Q000378:metabolism; D007610:Kallikreins / Q000096:biosynthesis* / Q000235:genetics*; D007985:Leydig Cells / Q000378:metabolism*; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011994:Recombinant Proteins / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013379:Substrate Specificity; D014018:Tissue Distribution
D002364:Caseins; D018076:DNA, Complementary; D005353:Fibronectins; D018972:Insulin-Like Growth Factor Binding Protein 3; D011994:Recombinant Proteins; D001224:Aspartic Acid; D005780:Gelatin; D007610:Kallikreins; C418971:Klk27 protein, mouse; D002918:Chymotrypsin; D005998:Glycine