Back to Search
Start Over
Expression cloning of a receptor for C5a anaphylatoxin on differentiated HL-60 cells
- Source :
- Biochemistry. 30(12)
- Publication Year :
- 1991
-
Abstract
- 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%.
- Subjects :
- Anaphylatoxins
Cellular differentiation
Molecular Sequence Data
Complement C5a
Biology
Molecular cloning
Transfection
Biochemistry
C5a receptor
Cell Line
Radioligand Assay
Dogs
Complementary DNA
Sequence Homology, Nucleic Acid
Animals
Humans
Amino Acid Sequence
RNA, Messenger
Cloning, Molecular
Receptor
Receptor, Anaphylatoxin C5a
COS cells
Anaphylatoxin receptors
Cell Differentiation
DNA
Blotting, Northern
Molecular biology
Receptors, Complement
Cross-Linking Reagents
Gene Expression Regulation
Expression cloning
Sequence Alignment
Subjects
Details
- ISSN :
- 00062960
- Volume :
- 30
- Issue :
- 12
- Database :
- OpenAIRE
- Journal :
- Biochemistry
- Accession number :
- edsair.doi.dedup.....d25e359edc03777af6ac37e5449ab5dc