Purification of a monocyte chemotactic factor secreted by nonhuman primate vascular cells in culture

Biochemistry. 1988 May 31;27(11):4162-8. doi: 10.1021/bi00411a039.

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 microgram 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.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acids / analysis
  • Animals
  • Cells, Cultured
  • Chemotactic Factors / isolation & purification*
  • Chemotactic Factors / metabolism
  • Chromatography, Gel
  • Chromatography, High Pressure Liquid
  • Culture Media
  • Electrophoresis, Polyacrylamide Gel
  • Monocytes / physiology*
  • Muscle, Smooth, Vascular / analysis*
  • Papio
  • Proteins / analysis

Substances

  • Amino Acids
  • Chemotactic Factors
  • Culture Media
  • Proteins