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

Ichijo, Hidenori; Rönnstrand, Lars LU ; Miyagawa, K; Ohashi, H; Heldin, Carl-Henrik and Miyazono, Kohei (1991) In Journal of Biological Chemistry 266(33). p.22459-22464
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... (More)
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. (Less)
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@article{07dd3f7d-6fba-4996-9f68-0475d36db502,
  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.},
  author       = {Ichijo, Hidenori and Rönnstrand, Lars and Miyagawa, K and Ohashi, H and Heldin, Carl-Henrik and Miyazono, Kohei},
  issn         = {1083-351X},
  keyword      = {Cell Surface/isolation & purification/metabolism
Receptors,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/metabolism
Molecular Weight
Platelet-Derived Growth Factor/metabolism
Receptors,Affinity
Chromatography,Animals
Carrier Proteins/*isolation & purification/metabolism
Cell Membrane/metabolism
Chromatography,Platelet-Derived Growth Factor
Recombinant Proteins/metabolism
Swine
Transforming Growth Factor beta/*metabolism
Uterus/*metabolism},
  language     = {eng},
  number       = {33},
  pages        = {22459--22464},
  publisher    = {ASBMB},
  series       = {Journal of Biological Chemistry},
  title        = {Purification of transforming growth factor-beta 1 binding proteins from porcine uterus membranes},
  volume       = {266},
  year         = {1991},
}