Purification and biochemical properties of a human monocyte-derived growth factor

J. P. Singh, P. D. Bonin

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

A monocyte-derived growth factor (MDGF) that stimulates proliferation of fibroblast and smooth muscle cells was purified from mitogen-stimulated human peripheral blood lymphocyte conditioned medium by using anion-exchange, Bio-Sil TSK-250 HPLC gel-permeation chromatography, and NaDodSO4/PAGE. Purified MDGF exhibited acidic charge characteristics (pI 5.0) and migrated with an apparent M(r) of 40,000 ± 2000 in molecular sizing HPLC colums. Elution from NaDodSO4/polyacrylamide gels showed that the growth-promoting activity was associated with three or four protein bands. The highest molecular weight species representing the most intense silver-stained band corresponded to 42,000; the lowest molecular weight species was 33,000. MDGF activity was stable to treatment with acid (pH 2.0) or base (pH 10.0) and heating (100°C, 5 min) but was inactivated upon reduction with 2-mercaptoethanol. The acidic MDGF did not effectively compete with platelet-derived growth factor (PDGF) for receptor binding and was not inhibited by PDGF antibodies. Previous studies have suggested that fibroblast growth-stimulating activity of macrophages is largely due to their secretion of interleukin 1 and a PDGF-like molecule. Our purification and biochemical characterization studies reveal the occurrence of multiple forms of fibroblast growth-stimulating activity in human monocyte conditioned medium. The MDGF activity characterized here appears to be structurally and functionally distinct from the previously described fibroblast growth-promoting activities including interleukin 1, basic fibroblast growth factor, and PDGF.

Original languageEnglish (US)
Pages (from-to)6374-6378
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume85
Issue number17
DOIs
StatePublished - Jan 1 1988

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Monocytes
Intercellular Signaling Peptides and Proteins
Platelet-Derived Growth Factor
Fibroblasts
Conditioned Culture Medium
Interleukin-1
Molecular Weight
High Pressure Liquid Chromatography
Platelet-Derived Growth Factor Receptors
Mercaptoethanol
Fibroblast Growth Factor 2
Growth
Mitogens
Silver
Human Activities
Heating
Smooth Muscle Myocytes
Gel Chromatography
Anions
Macrophages

ASJC Scopus subject areas

  • General

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Purification and biochemical properties of a human monocyte-derived growth factor. / Singh, J. P.; Bonin, P. D.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 85, No. 17, 01.01.1988, p. 6374-6378.

Research output: Contribution to journalArticle

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