Purification and partial characterization of a receptor protein for mouse interferon /gamma/
A receptor protein for mouse interferon /gamma/ has been purified from solubilized plasma membranes of the mouse monomyelocytic cell line WEHI-3. Sequential wheat germ agglutinin and ligand affinity chromatography of membranes extracted with octyl /beta/-D-glucopyranoside resulted in at least a 680-fold purification of the receptor, as measured by precipitating it in association with liposomes composed of phosphatidylcholine. The purified receptor bound /sup 125/I-labeled recombinant mouse interferon /gamma/ (rMuIFN-/gamma/) with a K/sub d/ of 10 nM, a value comparable to that obtained with isolated membranes, PAGE analysis of radiolabeled (with either /sup 35/S or /sup 125/I) receptor preparations consistently revealed a major band of 95 kDa. This species was degraged with time to smaller fragments, GR-20, a monoclonal antibody against the receptor, completely inhibited specific binding of /sup 125/I-labeled rMuIFN-/gamma/ to WEHI-3 cells, blocked the induction of priming by rMuIFN-/gamma/ of macrophage-mediated tumor cell killing, removed binding activity for /sup 125/I-labeled rMuIFN-/gamma/ from solubilized membranes, and immunoprecipitated a single 95-kDa protein from the extract of surface labeled (/sup 125/I) WEHI-3 cells. Cross-linking of /sup 125/I-labeled rMuIFN-/gamma/ to its receptor yielded a complex of 125 /plus minus/ 5 kDa, consistent with the binding of the dimeric form of mouse interferon /gamma/ (32 kDa) to a membrane protein of 95 kDa. These data suggest that the receptor for mouse interferon /gamma/ is a glycoprotein of 95 kDa.
- Research Organization:
- Univ. of Florida, Gainesville (USA)
- OSTI ID:
- 5702671
- Journal Information:
- Proc. Natl. Acad. Sci. USA; (United States), Journal Name: Proc. Natl. Acad. Sci. USA; (United States) Vol. 85:17; ISSN PNASA
- Country of Publication:
- United States
- Language:
- English
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59 BASIC BIOLOGICAL SCIENCES
ALKALI METAL COMPOUNDS
ANTIBODIES
AUTORADIOGRAPHY
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CARBOHYDRATES
CELL CONSTITUENTS
CELL MEMBRANES
CHEMICAL REACTIONS
CROSS-LINKING
DAYS LIVING RADIOISOTOPES
ELECTRON CAPTURE RADIOISOTOPES
ELECTROPHORESIS
EVEN-ODD NUCLEI
GLUCOPROTEINS
GLYCOPROTEINS
GROWTH FACTORS
HALIDES
HALOGEN COMPOUNDS
INORGANIC PHOSPHORS
INTERFERON
INTERMEDIATE MASS NUCLEI
IODIDES
IODINE 125
IODINE COMPOUNDS
IODINE ISOTOPES
ISOTOPES
LABELLED COMPOUNDS
LIGHT NUCLEI
LYMPHOKINES
MEMBRANE PROTEINS
MEMBRANES
MITOGENS
MOLECULAR STRUCTURE
MONOCLONAL ANTIBODIES
NUCLEI
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
PHOSPHORS
POLYMERIZATION
PROTEINS
PURIFICATION
RADIOISOTOPES
RECEPTORS
SACCHARIDES
SODIUM COMPOUNDS
SODIUM IODIDES
SULFUR 35
SULFUR ISOTOPES