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Author Gupta, S. L.
Source United States Department of Energy Office of Scientific and Technical Information
Content type Text
Language English
Subject Keyword BASIC BIOLOGICAL SCIENCES ♦ INTERFERON ♦ CHEMICAL BONDS ♦ RECEPTORS ♦ CROSS-LINKING ♦ GENE RECOMBINATION ♦ IODINE 125 ♦ LABELLED COMPOUNDS ♦ MAN ♦ TRACER TECHNIQUES ♦ ANIMALS ♦ BETA DECAY RADIOISOTOPES ♦ CHEMICAL REACTIONS ♦ DAYS LIVING RADIOISOTOPES ♦ ELECTRON CAPTURE RADIOISOTOPES ♦ INTERMEDIATE MASS NUCLEI ♦ IODINE ISOTOPES ♦ ISOTOPE APPLICATIONS ♦ ISOTOPES ♦ MAMMALS ♦ NUCLEI ♦ ODD-EVEN NUCLEI ♦ POLYMERIZATION ♦ PRIMATES ♦ RADIOISOTOPES ♦ VERTEBRATES ♦ Biochemistry- Tracer Techniques
Abstract Purified recombinant human ..gamma.. interferon (HuIFN-..gamma..), labeled with /sup 125/I (/sup 125/I-HuIFN-..gamma..), was used to study receptors for HuIFN-..gamma.. on human WISH cells. /sup 125/I-HuIFN-..gamma.. was bound to WISH cells, and this binding was displaced by unlabeled HuIFN-..gamma.. but not by unlabeled recombinant HuIFN-..cap alpha../sub 2/ or (Ser/sup 17/)HuIFN-..beta.. (HuIFN-..beta.. with serine substituted for cysteine at position 17), indicating the presence of specific binding sites for HuIFN-..gamma... The cell-bound /sup 125/I-HuINF-..gamma.. was crosslinked with disuccinimidyl suberate or ethylene glycol bis(succinimidyl succinate), which yielded a complex of M/sub r/ approx. = 105,000 +/- 5000 as analyzed by NaDodSO/sub 4//PAGE. The formation of this complex was prevented by preincubation of cells with unlabeled HuIFN-..gamma.. but not with HuIFN-..cap alpha../sub 2/ or (Ser/sup 17/)HuIFN-..beta.., indicating that HuIFN-..gamma.. binds to specific receptor molecule and that HuIFN-..cap alpha../sub 2/ or HuIFN-..beta.. do not interact with this receptor. Experiments were carried out with /sup 125/I-labeled recombinant (Ser/sup 17/)HuIFN-..beta.. (/sup 125/I-(Ser/sup 17/)HuIFN-..beta..) to verify this conclusion. Binding and crosslinking of /sup 125/I-(Ser/sup 17/)HuIFN-..beta.. to human WISH cells and Daudi cells yielded a complex of M/sub r/ 4z 150,000 similar to that obtained with /sup 125/I-HuIFN-..cap alpha.. as described earlier. The formation of this M/sub r/ 150,000 complex with /sup 125/I-(Ser/sup 17/)HuIFN-..beta.. was displaced by unlabeled (Ser/sup 17/)HuIFN-..beta.. and by HuINF-..cap alpha.. but not by HuIFN-..gamma.., indicating that (Ser/sup 17/)HuIFN-..beta.. binds to the same receptor as does HuIFN-..cap alpha../sub 2/, identified earlier, and that HuIFN-..gamma.. does not compete with /sup 125/I-(Ser/sup 17/)HuIFN-..beta.. for this receptor. 34 references, 7 figures.
Educational Use Research
Learning Resource Type Article
Publisher Date 1984-08-01
Publisher Place United States
Volume Number 81
Issue Number 16
Organization Cornell Univ., New York, NY


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