|
|
 Previous Article | Table of Contents | Next Article 
Purification and properties of bacterially synthesized human
granulocyte-macrophage colony stimulating factor
AW Burgess, CG Begley, GR Johnson, AF Lopez, DJ Williamson, JJ Mermod, RJ Simpson, A Schmitz and JF DeLamarter
Human granulocyte-macrophage colony stimulating factor (GM-CSF) has been
synthesized in high yield using a temperature inducible plasmid in
Escherichia coli. The human GM-CSF is readily isolated from the bacterial
proteins because of its differential solubility and chromatographic
properties. The bacterially synthesized form of the human GM-CSF contains
an extra methionine residue at position 1, but otherwise it is identical to
the polypeptide predicted from the cDNA sequence. The specific activity of
2.9 X 10(7) units/mg of protein for purified bacterially synthesized human
GM-CSF indicates that despite the lack of glycosylation, the molecule is
substantially in its native conformation. This molecule stimulated the same
number and type of both seven- and 14-day human bone marrow colonies as the
CSF alpha preparation from human placental conditioned medium. Human GM-CSF
had no activity on murine bone marrow or murine leukemic cells. There was
no detectable, direct stimulation of adult human erythroid burst forming
units (BFU-E) by the bacterially synthesized human GM-CSF. Although impure
preparations containing native human GM-CSF (eg, human placental
conditioned medium) stimulated the formation of mixed colonies, even in the
presence of erythropoietin, the bacterially synthesized human GM-CSF failed
to stimulate the formation of mixed colonies from adult human bone marrow
cells. The bacterially synthesized human GM-CSF increased
N-formyl-methionyl-leucyl- phenylalanine (FMLP)-induced superoxide
production and lysozyme secretion. Antibody-dependent cytotoxicity and
phagocytosis by human neutrophils was stimulated by the bacterially
synthesized human GM-CSF and eosinophils were also activated in the
antibody-dependent cytotoxicity assay.
Volume 69,
Issue 1,
pp. 43-51,
01/01/1987
Copyright © 1987 by The American Society of Hematology
 CiteULike  Connotea  Del.icio.us  Digg  Reddit  Technorati What's this?
This article has been cited by other articles:
|
|
|
|
 
A. Sebollela, T. C. Cagliari, G. S. C. S. Limaverde, A. Chapeaurouge, M. H. F. Sorgine, T. Coelho-Sampaio, C. H. I. Ramos, and S. T. Ferreira
Heparin-binding Sites in Granulocyte-Macrophage Colony-stimulating Factor: LOCALIZATION AND REGULATION BY HISTIDINE IONIZATION
J. Biol. Chem.,
September 9, 2005;
280(36):
31949 - 31956.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. Wadhwa, A.-L. H. Skog, C. Bird, P. Ragnhammar, M. Lilljefors, R. Gaines-Das, H. Mellstedt, and R. Thorpe
Immunogenicity of Granulocyte-Macrophage Colony-stimulating Factor (GM-CSF) Products in Patients Undergoing Combination Therapy with GM-CSF
Clin. Cancer Res.,
June 1, 1999;
5(6):
1353 - 1361.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. O. Armitage
Emerging Applications of Recombinant Human Granulocyte-Macrophage Colony-Stimulating Factor
Blood,
December 15, 1998;
92(12):
4491 - 4508.
[Full Text]
[PDF]
|
|
|
|
|
|
|
C.G. Begley, R. Basser, R. Mansfield, B. Thomson, W.R.L. Parker, J. Layton, B. To, J. Cebon, W.P. Sheridan, R.M. Fox, et al.
Enhanced Levels and Enhanced Clonogenic Capacity of Blood Progenitor Cells Following Administration of Stem Cell Factor Plus Granulocyte Colony-Stimulating Factor to Humans
Blood,
November 1, 1997;
90(9):
3378 - 3389.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
K Diederichs, T Boone, and P. Karplus
Novel fold and putative receptor binding site of granulocyte-macrophage colony-stimulating factor
Science,
December 20, 1991;
254(5039):
1779 - 1782.
[Abstract]
[PDF]
|
|
|
|
|
