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Purification of the platelet-derived growth factor receptor by using an anti-phosphotyrosine antibody

Journal Article · · Proc. Natl. Acad. Sci. U.S.A.; (United States)
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The platelet-derived growth factor (PDGF) receptor is a 180-kDa membrane glycoprotein. A protein of identical size, lectin affinity, and isoelectric point has been identified as a major substrate for PDGF-activated tyrosine kinase in stimulated 3T3 cells. The authors have purified this tyrosine-phosphorylated protein to homogeneity by using anti-phosphotyrosine immunoaffinity and lectin affinity steps. Demonstration that this purified tyrosine phosphoprotein is the PDGF receptor necessitated development of an assay capable of identifying specific SVI-labeled PDGF binding activity in soluble receptor preparations. Precipitated binding sites display affinity and kinetic characteristics of PDGF receptors in cells and membranes. Preparations of the 180-kDa phosphoprotein that are > 90% homogeneous by silver stain and by (TVS)methionine protein autoradiography have specific high affinity SVI-labeled PDGF binding sites. These data demonstrate that the 180-kDa substrate of the PDGF-stimulated tyrosine kinase is the PDGF receptor. Furthermore, these methods provide a means of purifying this and other tyrosine kinase substrates from growth factor-stimulated cells.

Research Organization:
Univ. of California, San Francisco
OSTI ID:
6085609
Journal Information:
Proc. Natl. Acad. Sci. U.S.A.; (United States), Journal Name: Proc. Natl. Acad. Sci. U.S.A.; (United States) Vol. 82:9; ISSN PNASA
Country of Publication:
United States
Language:
English

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Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AMINOTRANSFERASES
ANIMAL CELLS
ANTIBODIES
AUTORADIOGRAPHY
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOCHEMISTRY
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BLOOD PLATELETS
BODY FLUIDS
CARBOHYDRATES
CELL CONSTITUENTS
CELL MEMBRANES
CHEMISTRY
CONNECTIVE TISSUE CELLS
DAYS LIVING RADIOISOTOPES
ELECTRON CAPTURE RADIOISOTOPES
ENZYMES
EVEN-ODD NUCLEI
FIBROBLASTS
GLUCOPROTEINS
INTERMEDIATE MASS NUCLEI
IODINE 125
IODINE ISOTOPES
ISOTOPE APPLICATIONS
ISOTOPES
LIGHT NUCLEI
MATERIALS
MEMBRANES
NITROGEN TRANSFERASES
NUCLEI
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
PROTEINS
RADIOISOTOPES
RADIORECEPTOR ASSAY
RECEPTORS
SACCHARIDES
SOMATIC CELLS
SULFUR 35
SULFUR ISOTOPES
TRACER TECHNIQUES
TRANSFERASES