Purification of transforming growth factor-beta 1 binding proteins from porcine uterus membranes

J Biol Chem. 1991 Nov 25;266(33):22459-64.

Abstract

We have identified several transforming growth factor-beta 1 (TGF-beta 1) binding proteins in solubilized and glycoprotein-enriched porcine uterus membrane fractions by affinity cross-linking and in-gel ligand binding using 125I-labeled TGF-beta 1. By a ligand affinity chromatography using a column of immobilized recombinant TGF-beta 1, four components of apparent molecular weights 160,000, 80,000, 50,000, and 40,000 under reducing conditions were eluted at a pH of 3.5; the 160-,80-, and 40-kDa components were demonstrated to bind TGF-beta 1 specifically by the 125I-TGF-beta 1 binding assays. Further purification was performed by gel chromatography using a Superose 12 column eluted in 70% formic acid. The 40-kDa component was purified to an apparently homogenous form, whereas the 160-kDa component eluted in a broad peak overlapping the peak of the 80-kDa component. It remains to be elucidated whether these TGF-beta 1 binding proteins are related to cell surface receptors for TGF-beta s.

MeSH terms

  • Animals
  • Carrier Proteins / isolation & purification*
  • Carrier Proteins / metabolism
  • Cell Membrane / metabolism
  • Chromatography, Affinity
  • Chromatography, Gel
  • Chromatography, High Pressure Liquid
  • Chromatography, Ion Exchange
  • Electrophoresis, Polyacrylamide Gel
  • Female
  • Intracellular Signaling Peptides and Proteins*
  • Latent TGF-beta Binding Proteins
  • Membrane Glycoproteins / isolation & purification*
  • Membrane Glycoproteins / metabolism
  • Molecular Weight
  • Platelet-Derived Growth Factor / metabolism
  • Receptors, Cell Surface / isolation & purification
  • Receptors, Cell Surface / metabolism
  • Receptors, Platelet-Derived Growth Factor
  • Recombinant Proteins / metabolism
  • Swine
  • Transforming Growth Factor beta / metabolism*
  • Uterus / metabolism*

Substances

  • Carrier Proteins
  • Intracellular Signaling Peptides and Proteins
  • Latent TGF-beta Binding Proteins
  • Membrane Glycoproteins
  • Platelet-Derived Growth Factor
  • Receptors, Cell Surface
  • Recombinant Proteins
  • Transforming Growth Factor beta
  • Receptors, Platelet-Derived Growth Factor