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Purification and Partial Amino Acid Sequence of Asialo Murine Granulocyte--Macrophage Colony Stimulating Factor

Sparrow, Lindsay G. and Metcalf, Donald and Hunkapiller, Michael W. and Hood, Leroy E. and Burgess, Anthony W. (1985) Purification and Partial Amino Acid Sequence of Asialo Murine Granulocyte--Macrophage Colony Stimulating Factor. Proceedings of the National Academy of Sciences of the United States of America, 82 (2). pp. 292-296. ISSN 0027-8424. http://resolver.caltech.edu/CaltechAUTHORS:SPApnas85

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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.


Item Type:Article
Additional Information:Copyright © 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.
Subject Keywords:neuraminidase treatment; HPLC; sequence microanalysis; mouse lung-conditioned medium
Record Number:CaltechAUTHORS:SPApnas85
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:SPApnas85
Alternative URL:http://www.pnas.org/cgi/content/abstract/82/2/292
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:5726
Collection:CaltechAUTHORS
Deposited By: Archive Administrator
Deposited On:30 Oct 2006
Last Modified:26 Dec 2012 09:14

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