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Title: 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

Abstract

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 proposemore » to designate the low-affinity GM-CSF receptor and the KH97 protein as the {alpha} and {beta} subunits of the GM-CSF receptor, respectively.« less

Authors:
; ; ;  [1]; ;  [2]
  1. DNAX Research Inst. of Molecular and Cellular Biology, Palo Alto, CA (United States)
  2. Univ. of Tokyo (Japan)
Publication Date:
OSTI Identifier:
5044390
Resource Type:
Journal Article
Journal Name:
Proceedings of the National Academy of Sciences of the United States of America; (United States)
Additional Journal Information:
Journal Volume: 87:24; Journal ID: ISSN 0027-8424
Country of Publication:
United States
Language:
English
Subject:
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

Citation Formats

Hayashida, Kazuhiro, Kitamura, Toshio, Gorman, D M, Miyajima, Atsushi, Arai, Kenichi, and Yokota, Takashi. 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. United States: N. p., 1990. Web. doi:10.1073/pnas.87.24.9655.
Hayashida, Kazuhiro, Kitamura, Toshio, Gorman, D M, Miyajima, Atsushi, Arai, Kenichi, & Yokota, Takashi. 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. United States. https://doi.org/10.1073/pnas.87.24.9655
Hayashida, Kazuhiro, Kitamura, Toshio, Gorman, D M, Miyajima, Atsushi, Arai, Kenichi, and Yokota, Takashi. 1990. "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". United States. https://doi.org/10.1073/pnas.87.24.9655.
@article{osti_5044390,
title = {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},
author = {Hayashida, Kazuhiro and Kitamura, Toshio and Gorman, D M and Miyajima, Atsushi and Arai, Kenichi and Yokota, Takashi},
abstractNote = {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.},
doi = {10.1073/pnas.87.24.9655},
url = {https://www.osti.gov/biblio/5044390}, journal = {Proceedings of the National Academy of Sciences of the United States of America; (United States)},
issn = {0027-8424},
number = ,
volume = 87:24,
place = {United States},
year = {Sat Dec 01 00:00:00 EST 1990},
month = {Sat Dec 01 00:00:00 EST 1990}
}