Receptors for human. gamma. interferon: binding and crosslinking of /sup 125/I-labeled recombinant human. gamma. interferon to receptors on WISH cells

Journal Article · Wed Aug 01 00:00:00 EDT 1984 · Proc. Natl. Acad. Sci. U.S.A.; (United States)

DOI:https://doi.org/10.1073/pnas.81.16.5160· OSTI ID:5809058

Sarkar, F H; Gupta, S L

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.

• Research Organization: Cornell Univ., New York, NY
• OSTI ID: 5809058
• Journal Information: Proc. Natl. Acad. Sci. U.S.A.; (United States), Vol. 81:16
• Country of Publication: United States
• Language: English

Related Subjects

59 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
550201* - Biochemistry- Tracer Techniques