Purification of a Monocyte Chemotactic Factor Secreted by Nonhuman Primate Vascular Cells in Culture

Anthony J. Valente, Dana T. Gravesl, Catherine E. Vialle-valentin, Ruby Delgado, Colin J. Schwartz

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211 Scopus citations

Abstract

A protein chemotactic for peripheral blood monocytes (SMC-CF) of potential importance in their recruitment to the arterial intima in atherogenesis was purified from serum-free medium conditioned by cultured baboon aortic medial smooth muscle cells. The purification of SMC-CF was monitored by a filter assay using human peripheral blood mononuclear cells and was achieved by batch separation on a cation-exchange gel followed by gel permeation chromatography, ion-exchange high-performance liquid chromatography (HPLC), and reversed-phase HPLC. The overall recovery was approximately 10% of the initial activity and yielded 0.5-1 μg of SMC-CF/L of conditioned medium. On analytical sodium dodecyl sulfate-polyacrylamide gel electrophoresis, SMC-CF migrated as a monomeric protein with an apparent molecular weight of 14 500. A dose-dependent relationship was observed between SMC-CF concentration and monocyte chemotactic activity, with maximal and half-maximal biologic activity being observed at approximately 5 and 0.1 nM, respectively. Cultured baboon aortic smooth muscle cells also express the genes for both the A and B polypeptide chains of platelet-derived growth factor, which has been reported to be chemotactic for blood monocytes and neutrophils [Deuel, T. F., Senior, R. M., Huang, J. S., & Griffin, G. L. (1982) J. Clin. Invest. 69, 1046–1049]. Amino acid composition analyses indicate that SMC-CF is not derived either from polypeptide chain of this growth factor or from certain potentially chemotactic connective tissue proteins.

Original languageEnglish (US)
Pages (from-to)4162-4168
Number of pages7
JournalBiochemistry
Volume27
Issue number11
DOIs
StatePublished - May 1 1988

ASJC Scopus subject areas

  • Biochemistry

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    Valente, A. J., Gravesl, D. T., Vialle-valentin, C. E., Delgado, R., & Schwartz, C. J. (1988). Purification of a Monocyte Chemotactic Factor Secreted by Nonhuman Primate Vascular Cells in Culture. Biochemistry, 27(11), 4162-4168. https://doi.org/10.1021/bi00411a039