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Isolation and partial amino acid sequence analysis of nerve growth factor from the guinea pig prostate
Author(s) -
Rubin Jeffrey S.,
Bradshaw Ralph A.
Publication year - 1981
Publication title -
journal of neuroscience research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.72
H-Index - 160
eISSN - 1097-4547
pISSN - 0360-4012
DOI - 10.1002/jnr.490060403
Subject(s) - nerve growth factor , guinea pig , size exclusion chromatography , biochemistry , arginine , peptide sequence , amino acid , microbiology and biotechnology , polyacrylamide gel electrophoresis , biology , chemistry , enzyme , endocrinology , receptor , gene
Nerve growth factor (NGF) has been purified from the guinea pig prostate using a modification of the Bocchini‐Angeletti method for isolating 2.5S NGF from mouse submaxillary glands. As with the mouse preparation, guinea pig prostate NGF appears to migrate as a high molecular weight entity at physiological pH. Following dissociation, NGF, active in neurite proliferation assays and similar in size to mouse 2.5S NGF, can be isolated by chromatography on a column of carboxymethyl‐cellulose at pH 4.8. Based on gel filtration, SDS‐polyacrylamide gel analysis, and amino‐terminal sequence studies, this material consists of two, noncovalently linked, identical polypeptide chains each with a molecular weight of about 13,000. The amino‐terminal third of the polypeptide chain is at least 90% identical to the corresponding region of the murine molecule, confirming the homology of the guinea pig prostate protein to NGFs obtained from different tissues in other species. However, in contrast to the mouse preparation, the putative high molecular weight form of guinea pig NGF does not contain a subunit with arginine esteropeptidase activity. Although there is an abundance of this enzymatic activity in the homogenate, it does not appear to be associated with the fractions containing NGF. This apparent difference in the mouse and guinea pig material is of interest because the mouse γ subunit, possessing the arginine esteropeptidase activity, has been alleged to participate in the processing of a precursor of the β NGF polypeptide chain.