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), Vol. 85:17
- Country of Publication:
- United States
- Language:
- English
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LABELLED COMPOUNDS
MOLECULAR STRUCTURE
MONOCLONAL ANTIBODIES
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SODIUM IODIDES
SULFUR 35
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ANTIBODIES
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CARBOHYDRATES
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EVEN-ODD NUCLEI
GLYCOPROTEINS
GROWTH FACTORS
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HALOGEN COMPOUNDS
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INTERMEDIATE MASS NUCLEI
IODIDES
IODINE COMPOUNDS
IODINE ISOTOPES
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LIGHT NUCLEI
LYMPHOKINES
MEMBRANE PROTEINS
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NUCLEI
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SODIUM COMPOUNDS
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550201* - Biochemistry- Tracer Techniques