Native and activated forms of alpha 2-macroglobulin increase expression of platelet-derived growth factor alpha-receptor in vascular smooth muscle cells. Evidence for autocrine transforming growth factor-beta activity

J Biol Chem. 1995 Dec 22;270(51):30741-8. doi: 10.1074/jbc.270.51.30741.

Abstract

Cellular response to platelet-derived growth factor AA (PDGF-AA) is mediated exclusively by the PDGF alpha-receptor. Vascular smooth muscle cells (VSMCs) in culture typically express very low levels of alpha-receptor. In this study, we demonstrate that the proteinase inhibitor and cytokine carrier alpha 2-macroglobulin (alpha 2M) increases rat VSMC PDGF alpha-receptor expression. PDGF alpha-receptor mRNA levels increased 3-fold by 6 h and were sustained at that level through 24 h in VSMCs treated with 280 nM methylamine-modified alpha 2M (alpha 2M-MA), a form of activated alpha 2M. PDGF beta-receptor mRNA levels were unchanged in the same time period. In 125I-PDGF-AA binding experiments, treatment of VSMCs with alpha 2M-MA increased the maximum binding capacity (Bmax) from 1.9 to 9.2 fmol/mg of cell protein without affecting binding affinity (KD approximately 80 pM). alpha 2M-MA also increased the VSMC response to PDGF-AA as determined by tyrosine phosphorylation of a 170-kDa band, corresponding in mass to the PDGF alpha-receptor. The native form of alpha 2M was comparable to alpha 2M-MA in its ability to increase PDGF-AA binding to VSMCs and tyrosine phosphorylation of the 170-kDa band. Recombinant and proteolytic alpha 2M derivatives were used to demonstrate that alpha 2M increases PDGF alpha-receptor expression by binding VSMC-secreted cytokine(s) and interrupting an autocrine loop that ordinarily suppresses alpha-receptor expression in these cells. Transforming growth factor-beta-neutralizing antibody mimicked the activity of alpha 2M, increasing the binding capacity of VSMCs for PDGF-AA. This study demonstrates that VSMC PDGF alpha-receptor expression and responsiveness to PDGF-AA are regulated by autocrine transforming growth factor-beta activity, potentially other autocrine growth factors, and alpha 2M.

Publication types

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

MeSH terms

  • Animals
  • Aorta / metabolism
  • Cell Division / drug effects
  • Cells, Cultured
  • DNA / biosynthesis
  • Fibroblast Growth Factor 2 / metabolism
  • Gene Expression / drug effects
  • Humans
  • Kinetics
  • Muscle, Smooth, Vascular / cytology
  • Muscle, Smooth, Vascular / drug effects
  • Muscle, Smooth, Vascular / metabolism*
  • Platelet-Derived Growth Factor / metabolism*
  • RNA, Messenger / analysis
  • RNA, Messenger / biosynthesis
  • Rats
  • Rats, Sprague-Dawley
  • Receptor, Platelet-Derived Growth Factor alpha
  • Receptors, Platelet-Derived Growth Factor / biosynthesis*
  • Structure-Activity Relationship
  • Up-Regulation
  • alpha-Macroglobulins / pharmacology*

Substances

  • Platelet-Derived Growth Factor
  • RNA, Messenger
  • alpha-Macroglobulins
  • platelet-derived growth factor A
  • Fibroblast Growth Factor 2
  • DNA
  • Receptor, Platelet-Derived Growth Factor alpha
  • Receptors, Platelet-Derived Growth Factor