Expression cloning of a receptor for C5a anaphylatoxin on differentiated HL-60 cells

Journal Article · Tue Mar 26 04:00:00 EST 1991 · Biochemistry; (United States)

DOI:https://doi.org/10.1021/bi00226a002· OSTI ID:5510893

Boulay, F; Mery, L; Tardif, M; Brouchon, L; Vignais, P ^[1]

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1. Centre d'Etudes Nucleaires, Grenoble (France)

A cDNA clone encoding the human C5a anaphylatoxin receptor has been isolated by expression cloning from a CDM8 expression library prepared from mRNA of human myeloid HL-60 cells differentiated to the granulocyte phenotype with dibutyryladenosine cyclic monophosphate. The cDNA clone was able to transfer to COS-7 cells the capacity to specifically bind iodinated human recombinant C5a. The cDNA was 2.3 kb long, with an open reading frame encoding a 350-residue polypeptide. Cross-linking of iodinated C5a to the plasma membrane of transfected COS cells revealed a complex with an apparent molecular mass of 52-55 kDa, similar to that observed for the constitutively expressed receptor in differentiated HL-60 cells or human neutrophils. Although differentiated HL-60 cells display a single class of binding sites, with a dissociation constant of approximately 800-900 pM, the C5a-R cDNA, expressed in COS cells, generates both high-affinity (1.7 nM) and low-affinity (20-25 nM) receptors. Sequence comparison established that the degree of sequence identity between the C5a receptor and the N-formlypeptide receptor is 34%.

• OSTI ID: 5510893
• Journal Information: Biochemistry; (United States), Journal Name: Biochemistry; (United States) Vol. 30:12; ISSN 0006-2960; ISSN BICHA
• Country of Publication: United States
• Language: English

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59 BASIC BIOLOGICAL SCIENCES
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DNA SEQUENCING
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NUCLEIC ACIDS
ODD-EVEN NUCLEI
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RECEPTORS
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RNA
STRUCTURAL CHEMICAL ANALYSIS
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