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

Journal Article · Wed May 01 00:00:00 EDT 1985 · Proc. Natl. Acad. Sci. U.S.A.; (United States)

DOI:https://doi.org/10.1073/pnas.82.9.2684· OSTI ID:6085609

Daniel, T O; Tremble, P M; Frackelton, Jr, A R; Williams, L T

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, CA (United States)
• OSTI ID: 6085609
• Journal Information: Proc. Natl. Acad. Sci. U.S.A.; (United States), Vol. 82:9
• Country of Publication: United States
• Language: English

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59 BASIC BIOLOGICAL SCIENCES
AMINOTRANSFERASES
BIOCHEMISTRY
CELL MEMBRANES
AUTORADIOGRAPHY
GLUCOPROTEINS
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ANTIBODIES
BLOOD PLATELETS
FIBROBLASTS
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SULFUR 35
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BLOOD
BLOOD CELLS
BODY FLUIDS
CARBOHYDRATES
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DAYS LIVING RADIOISOTOPES
ELECTRON CAPTURE RADIOISOTOPES
ENZYMES
EVEN-ODD NUCLEI
INTERMEDIATE MASS NUCLEI
IODINE ISOTOPES
ISOTOPE APPLICATIONS
ISOTOPES
LIGHT NUCLEI
MATERIALS
MEMBRANES
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NUCLEI
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
PROTEINS
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550201* - Biochemistry- Tracer Techniques