Purification and Partial Amino Acid Sequence of Asialo Murine Granulocyte--Macrophage Colony Stimulating Factor
Abstract
A procedure utilizing reversed-phase high-performance liquid chromatography is described for the purification of asialo granulocyte--macrophage colony stimulating factor (asialo-GM-CSF) from mouse lung-conditioned medium. In the purification, the partially purified factor was treated with neuraminidase to reduce charge heterogeneity due to variable degrees of sialation. Three active forms of the asialo factor were separated by the final reversed-phase liquid chromatography step. These each gave a single major band and several minor bands on polyacrylamide gel electrophoresis and had similar amino acid compositions. The specific activity of purified murine asialo-GM-CSF was approximately 8 x 109 colonies per mg of protein. Amino acid sequence determination of the major form gave a single amino-terminal sequence, which has been used to develop oligonucleotide probes for the isolation of two cDNA clones encoding GM-CSF. The nucleotide sequence of these two clones gave a deduced amino acid sequence almost identical with that determined for the amino terminus of asialo-GM-CSF and an amino acid composition very similar to that for asialo-GM-CSF.
Additional Information
© 1985 by the National Academy of Sciences Communicated by G. J. V. Nossal, September 6, 1984 This work was supported in part by grants to D.M. from the Carden Fellowship Fund of the Anti-Cancer Council of Victoria and the National Institutes of Health (Bethesda, MD) (Grant CA 22556). The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.Attached Files
Published - SPApnas85.pdf
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Additional details
- PMCID
- PMC397023
- Eprint ID
- 5726
- Resolver ID
- CaltechAUTHORS:SPApnas85.961
- Anti-Cancer Council of Victoria
- CA 22556
- NIH
- Created
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2006-10-30Created from EPrint's datestamp field
- Updated
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2022-10-05Created from EPrint's last_modified field