Identification of autoproteolytic cleavage site in the Asp-49 mutant subtilisin J by site-directed mutagenesis.
Ser-49 located at the exposed surface loop of subtilisin J was replaced with Asp and Arg. Proteinase activity of the Asp-49 mutant was similar to that of wild-type, but the Arg mutant was inactivated. At 37 degrees C, mature subtilisin J protein of the Asp-49 mutant rapidly degraded, and specific breakdown products were accumulated. These proteins were analyzed by SDS-PAGE, and the N-terminal sequences were determined for the mature and deleted protein. We identified the autoproteolytic cleavage site in the mature Asp-49 mutant protein from sequencing data.
Jang
J S
JS
Department of Agricultural Chemistry, Korea University, Seoul.
Park
D K
DK
Chun
M
M
Byun
S M
SM
D000595:Amino Acid Sequence; D001120:Arginine; D001224:Aspartic Acid; D001407:Bacillus / Q000235:genetics*; D001665:Binding Sites; D004795:Enzyme Stability; D008970:Molecular Weight; D016297:Mutagenesis, Site-Directed; D011506:Proteins / Q000378:metabolism; D012694:Serine; D013381:Subtilisins / Q000737:chemistry / Q000235:genetics*
D011506:Proteins; C077349:subtilisin J; D001224:Aspartic Acid; D012694:Serine; D001120:Arginine; D013381:Subtilisins