The isolation and partial characterization of precursor forms of ostrich carboxypeptidase

Noxolo T. Mkwetshana, Ryno J. Naudé, Willem Oelofsen, Takako Naganuma, Koji Muramoto

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    2 Citations (Scopus)

    Abstract

    Ostrich carboxypeptidases A and B were recently purified and characterized. The aim of this study was to isolate and purify, and partially characterize in terms of molecular weight, pI, amino acid composition and N-terminal sequencing, the precursor forms of carboxypeptidases from the ostrich pancreas. Inhibition studies with soybean trypsin inhibitor and activation studies with three proteases (bovine trypsin, bovine chymotrypsin and porcine elastase) were performed on crude ostrich acetone powder and the carboxypeptidase A and B activities were determined. SDS-PAGE was carried out after every incubation to monitor the rate and degree of conversion of a M(r) 66K component to procarboxypeptidase and carboxypeptidase A and B. The precursor forms were purified by Toyopearl Super Q and Pharmacia Mono Q chromatography. All three proteases converted the M(r) 66K component to procarboxypeptidases and carboxypeptidases over a set time interval, with carboxypeptidase A and B activities being detected in the acetone powder. Chymotrypsin was the preferred protease since it exhibited a more controlled activation of the procarboxypeptidases. The amino acid composition of procarboxypeptidase A revealed 525 residues. The N-terminal sequence of procarboxypeptidase A showed considerable homology when compared with several other mammalian sequences. M(r) and pI values of 52K and 5.23 were obtained for procarboxypeptidase A, respectively, This study indicated that ostrich procarboxypeptidase A is closely related to other mammalian procarboxypeptidase A molecules in terms of physicochemical properties.

    Original languageEnglish
    Pages (from-to)331-343
    Number of pages13
    JournalInternational Journal of Biochemistry and Cell Biology
    Volume31
    Issue number2
    DOIs
    Publication statusPublished - 1999 Feb 1

    Keywords

    • Carboxypeptidase
    • Ostrich
    • Pancreas
    • Precursors

    ASJC Scopus subject areas

    • Biochemistry
    • Cell Biology

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