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Binding of /sup 125/I-labeled recombinant beta interferon (IFN-beta Ser17) to human cells

Journal Article · · Mol. Cell. Biol.; (United States)
; ;

The authors investigated the binding of /sup 125/I-labeled beta interferon (IFN-beta Ser17), a nonglycosylated recombinant human fibroblast interferon in which cysteine at position 17 is replaced by serine by site-specific mutagenesis. An optimized chloramine T radiolabeling method produced a highly labeled, fully active /sup 125/I-IFN suitable for these studies. Unlike the case with the chloramine T method, incorporation of a single mole of Bolton-Hunter reagent into a mole of IFN-beta Ser17 led to nearly complete loss of biological activity. /sup 125/I-IFN-beta Ser17, prepared by the chloramine T method, bound specifically to human lymphoblastoid cells (Daudi) with a dissociation constant of 0.24 nM. The number of binding sites per cell was 4,000. In competition assays, unlabeled beta interferons (native, recombinant IFN-beta Cys17, and various preparations of IFN-beta Ser17) equally displaced labeled IFN-beta Ser17 on Daudi cells. Recombinant IFN-alpha-1 displaced /sup 125/I-IFN-beta binding to Daudi cells less efficiently than did unlabeled native or recombinant beta interferon. However, at the concentrations tested, native gamma interferon showed no competition with /sup 125/I-IFN. The results indicate that IFN-beta Ser17 and native IFN-beta posses similar binding properties.

Research Organization:
Cetus Corp., Emeryville, CA
OSTI ID:
5757350
Journal Information:
Mol. Cell. Biol.; (United States), Journal Name: Mol. Cell. Biol.; (United States) Vol. 4:12; ISSN MCEBD
Country of Publication:
United States
Language:
English

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Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AMINES
AMINO ACIDS
ANIMAL CELLS
BETA DECAY RADIOISOTOPES
BIOCHEMICAL REACTION KINETICS
CARBOXYLIC ACIDS
CHEMICAL BONDS
CHLORAMINES
CONNECTIVE TISSUE CELLS
CYSTEINE
DAYS LIVING RADIOISOTOPES
ELECTRON CAPTURE RADIOISOTOPES
FIBROBLASTS
HYDROXY ACIDS
INTERFERON
INTERMEDIATE MASS NUCLEI
IODINE 125
IODINE ISOTOPES
ISOTOPE APPLICATIONS
ISOTOPES
KINETICS
LABELLED COMPOUNDS
MUTAGENESIS
NUCLEI
ODD-EVEN NUCLEI
ORGANIC ACIDS
ORGANIC CHLORINE COMPOUNDS
ORGANIC COMPOUNDS
ORGANIC HALOGEN COMPOUNDS
ORGANIC SULFUR COMPOUNDS
RADIOISOTOPES
REACTION KINETICS
SERINE
SOMATIC CELLS
THIOLS
TRACER TECHNIQUES