Two soluble forms of bovine carboxypeptidase H have different NH2-terminal sequences.
Carboxypeptidase H is an important enzyme in the biosynthesis of many peptide hormones. Development of a rapid isolation procedure led to the purification of two soluble forms from acidic extracts of bovine pituitary glands. These two forms differed in apparent molecular size (56 and 53 kDa). Both forms were found in the anterior lobe while only the 53-kDa form was found in posterior lobe. Digestion with N-glycosidase F demonstrated that these two forms are not due to alternative glycosylation of a common polypeptide core. Both forms bind antibodies raised against a COOH-terminal peptide of the full-length protein showing that the difference between them is not due to proteolysis at the COOH terminus. These results also argue against the idea that proteolysis of COOH-terminal domains converts the membrane-associated form of this protein into a soluble form. NH2-terminal sequence analysis demonstrated different NH2 termini. The NH2-terminal sequence of the 56-kDa form begins at the site predicted for signal peptide cleavage. Ion-exchange chromatography resolved the 56-kDa form from the 53-kDa form. The two forms were catalytically active with very similar properties. These results show that bovine carboxypeptidase H can be posttranslationally processed at alternative sites and provide evidence against the idea of a prosequence that must be removed before enzyme activity can be expressed.
Department of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, Missouri 63110.
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D000595:Amino Acid Sequence; D000645:Ammonium Sulfate; D000818:Animals; D043423:Carboxypeptidase H; D002268:Carboxypeptidases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D002417:Cattle; D002846:Chromatography, Affinity; D003600:Cytosol / Q000201:enzymology; D007527:Isoenzymes / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D007700:Kinetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010902:Pituitary Gland / Q000201:enzymology*; D012689:Sequence Homology, Nucleic Acid
D007527:Isoenzymes; D002268:Carboxypeptidases; D043423:Carboxypeptidase H; D000645:Ammonium Sulfate