Expression cloning of the human C3a anaphylatoxin receptor (C3aR) from differentiated U-937 cells

被引:148
作者
Crass, T [1 ]
Raffetseder, U [1 ]
Martin, U [1 ]
Grove, M [1 ]
Klos, A [1 ]
Kohl, J [1 ]
Bautsch, W [1 ]
机构
[1] HANNOVER MED SCH, INST MED MICROBIOL, D-30625 HANNOVER, GERMANY
关键词
expression cloning; C3a receptor; C3a anaphylatoxin;
D O I
10.1002/eji.1830260840
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
A cDNA clone encoding the human C3a anaphylatoxin receptor (C3aR) was isolated from a pcDNAI/Amp expression library prepared from U-937 cells which had been differentiated with dibutyryl cAMP to a macrophage-like phenotype. The cDNA clone contained an insert of 4.3 kbp and was able to confer to transfected human HEK-293 cells the capacity to bind specifically iodinated human C3a. Chinese hamster ovary cells co-transfected with this cDNA clone and a G-protein alpha subunit (G alpha-16) became functionally responsive to C3a and a C3a analog synthetic peptide, as measured by increased phosphoinositide hydrolysis. As inferred from the cDNA sequence, the clone encodes a 482-residue polypeptide with seven hydrophobic membrane-spanning helices and a high homology to the human C5a and formyl-Met-Leu-Phe receptors, Uniquely among the family of G-protein coupled receptors, the C3aR contains an exceptionally large second extracellular loop of approximately 175 residues. Northern hybridizations revealed an approximately 2.3-kb transcript as the major and an additional similar to 3.9 kb-transcript as a minor transcription product of the C3aR. The C3aR appears to be widely expressed in different lymphoid tissues, as shown by Northern hybridizations, providing evidence for a central role of the C3a anaphylatoxin in inflammatory processes.
引用
收藏
页码:1944 / 1950
页数:7
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