The cleavage of protein kinase A by the kinase-splitting membranal proteinase is reproduced by meprin beta.
The Kinase-Splitting Membranal Proteinase (KSMP) is a metallo-endoproteinase that clips off the carboxyl terminus tail of the catalytic (C) subunit of protein kinase A to yield a truncated, catalytically inactive protein (C'). Here we report (a) a new procedure for the purification of KSMP, yielding a major protein band in SDS-polyacrylamide gel electrophoresis that correlates with the characteristic KSMP activity; (b) the sequence of tryptic peptides obtained from this band, suggesting an identity between this protein and meprin beta; (c) the immuno-detection by specific anti-peptide antibodies of both the alpha and the beta subunits of meprin in KSMP preparations; (d) the stable expression of meprin beta in a mammalian cell line (293) to establish a clone that constitutively expresses the full-length precursor of meprin beta; and (e) the optimalization of the proteolytic activation of this precursor to obtain an enzyme exhibiting the specific KSMP cleavage, suggesting that KSMP is either derived from, or identical with, the meprin beta gene product. It is hoped that these results will shed light on the possible physiological role of KSMP and the way it may affect protein kinase A-mediated processes.
Chestukhin
A
A
Department of Biological Regulation, The Weizmann Institute of Science, Rehovot 76100, Israel. lishalt@weizmann.weizmann.ac.il
Muradov
K
K
Litovchick
L
L
Shaltiel
S
S
D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism*; D010450:Endopeptidases / Q000302:isolation & purification / Q000378:metabolism*; D004789:Enzyme Activation; D004792:Enzyme Precursors / Q000378:metabolism; D006868:Hydrolysis; D008666:Metalloendopeptidases / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D014176:Protein Biosynthesis; D051381:Rats; D017386:Sequence Homology, Amino Acid; D014357:Trypsin / Q000378:metabolism
D004792:Enzyme Precursors; D017868:Cyclic AMP-Dependent Protein Kinases; D010450:Endopeptidases; D014357:Trypsin; D008666:Metalloendopeptidases; C095663:meprin B; C059784:kinase-splitting membranal proteinase