Molecular cloning of a second subunit of the receptor for human granulocyte - macrophage colony-stimulating factor (GM-CSF): Reconstitution of a high-affinity GM-CSF receptor

Journal Article · Sat Dec 01 00:00:00 EST 1990 · Proceedings of the National Academy of Sciences of the United States of America; (United States)

DOI:https://doi.org/10.1073/pnas.87.24.9655· OSTI ID:5044390

Hayashida, Kazuhiro; Kitamura, Toshio; Gorman, D M; Miyajima, Atsushi ^[1]; Arai, Kenichi; Yokota, Takashi ^[2]

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1. DNAX Research Inst. of Molecular and Cellular Biology, Palo Alto, CA (United States)
2. Univ. of Tokyo (Japan)

Using the mouse interleukin 3 (IL-3) receptor cDNA as a probe, the authors obtained a monologous cDNA (KH97) from a cDNA library of a human hemopoietic cell line, TF-1. The protein encoded by the KH97 cDNA has 56% amino acid sequence identity with the mouse IL-3 receptor and retains features common to the family of cytokine receptors. Fibroblasts transfected with the KH97 cDNA expressed a protein of 120 kDa but did not bind any human cytokines, including IL-3 and granulocyte - macrophage colony-stimulating factor (GM-CSF). Interestingly, cotransfection of cDNAs for KH97 and the low-affinity human GM-CSF receptor in fibroblasts resulted in formation of a high-affinity receptor for GM-CSF. The dissociation rate of GM-CSF from the reconstituted high-affinity receptor was slower than that from the low-affinity site, whereas the association rate was unchanged. Cross-linking of {sup 125}I-labeled GM-CSF to fibroblasts cotransfected with both cDNAs revealed the same cross-linking patterns as in TF-1 cells - i.e., two major proteins of 80 and 120 kDa which correspond to the low-affinity GM-CSF receptor and the KH97 protein, respectively. These results indicate that the high-affinity GM-CSF receptor is composed of at least two components in a manner analogous to the IL-2 receptor. They therefore propose to designate the low-affinity GM-CSF receptor and the KH97 protein as the {alpha} and {beta} subunits of the GM-CSF receptor, respectively.

• OSTI ID: 5044390
• Journal Information: Proceedings of the National Academy of Sciences of the United States of America; (United States), Vol. 87:24; ISSN 0027-8424
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
LYMPHOKINES
CROSS-LINKING
MEMBRANE PROTEINS
AMINO ACID SEQUENCE
RECEPTORS
RECOMBINANT DNA
DNA-CLONING
HEMATOPOIETIC SYSTEM
IODINE 125
MACROPHAGES
NEUTROPHILS
PHOSPHORUS 32
STEM CELLS
ANIMAL CELLS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BODY
BODY FLUIDS
CHEMICAL REACTIONS
CLONING
CONNECTIVE TISSUE CELLS
DAYS LIVING RADIOISOTOPES
DNA
DNA HYBRIDIZATION
ELECTRON CAPTURE RADIOISOTOPES
GROWTH FACTORS
HYBRIDIZATION
INTERMEDIATE MASS NUCLEI
INTERNAL CONVERSION RADIOISOTOPES
IODINE ISOTOPES
ISOTOPES
LEUKOCYTES
LIGHT NUCLEI
MATERIALS
MITOGENS
MOLECULAR STRUCTURE
NUCLEI
NUCLEIC ACIDS
ODD-EVEN NUCLEI
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
PHAGOCYTES
PHOSPHORUS ISOTOPES
POLYMERIZATION
PROTEINS
RADIOISOTOPES
SOMATIC CELLS
550201* - Biochemistry- Tracer Techniques