Purification, Characterization, and Partial Amino Acid Sequence of Nerve Growth Factor from Cobra Venom

Ruth A. Hogue-Angeletti, William A. Frazier, John W. Jacobs, Hugh D. Niall, Ralph A. Bradshaw

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59 Scopus citations

Abstract

The nerve growth factor (NGF) from Naja naja (cobra) venom has been purified and its structure compared to the NGF from mouse submaxillary gland. A two-step purification procedure has been devised, consisting of a gel filtration step in 1 M acetic acid followed by chromatography of the active pool on carboxymethylcellulose at pH 5. The molecular weight of the native protein was found to be 28000, and this value was reduced by approximately one-half under denaturing conditions. These values are comparable to those obtained for mouse 2.5S or βNGF. Tryptic peptide maps of S -[14C]carboxymethyl NGF gave the number of labeled peptides expected for a structure composed of two identical or very similar subunits. Thus, the quaternary structures of mouse and cobra NGF are the same. Cyanogen bromide (CNBr) treatment of Naja naja NGF produced three fragments, of which two were purified to homogeneity. These fragments and the whole protein were analyzed in the automated protein Sequencer. The amino-terminal CNBr fragment of the protein was also subjected to digestion by thermolysin and the resultant peptides were purified and characterized. These data plus those from the characterization of the tryptic peptides provided the basis of the construction of a tentative primary structure of Naja naja NGF which is approximately 60% identical with mouse NGF.

Original languageEnglish (US)
Pages (from-to)26-34
Number of pages9
JournalBiochemistry
Volume15
Issue number1
DOIs
StatePublished - Jan 1 1976

ASJC Scopus subject areas

  • Biochemistry

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    Hogue-Angeletti, R. A., Frazier, W. A., Jacobs, J. W., Niall, H. D., & Bradshaw, R. A. (1976). Purification, Characterization, and Partial Amino Acid Sequence of Nerve Growth Factor from Cobra Venom. Biochemistry, 15(1), 26-34. https://doi.org/10.1021/bi00646a005