Structure-function studies of the C3a-receptor: C-terminal serine and threonine residues which influence receptor internalization and signaling
(2003) In European Journal of Immunology 33(4). p.920-927- Abstract
- The anaphylatoxic peptide C3a is a pro-inflammatory mediator generated during complement activation, whose specific G protein coupled receptor is expressed on granulocytes, monocytes, mast cells, activated lymphocytes, and in the nervous tissue. We have generated RBL-2H3 cell clones stably expressing mutants of the human C3a-receptor (C3aR) with combined alanine (Ala) substitutions of ten C-terminal serine (Ser) or threonine (Thr) residues, which may represent putative phosphorylation sites to characterize their role in ligand-induced C3aR internalization and signaling. Ser475/479 and Thr480/481 as well as Ser449 seemed not to be involved in ligand-induced receptor internalization. Either directly or by a conformational change they even... (More)
- The anaphylatoxic peptide C3a is a pro-inflammatory mediator generated during complement activation, whose specific G protein coupled receptor is expressed on granulocytes, monocytes, mast cells, activated lymphocytes, and in the nervous tissue. We have generated RBL-2H3 cell clones stably expressing mutants of the human C3a-receptor (C3aR) with combined alanine (Ala) substitutions of ten C-terminal serine (Ser) or threonine (Thr) residues, which may represent putative phosphorylation sites to characterize their role in ligand-induced C3aR internalization and signaling. Ser475/479 and Thr480/481 as well as Ser449 seemed not to be involved in ligand-induced receptor internalization. Either directly or by a conformational change they even "inhibit" C3aR internalization. In contrast, mutants with Ala substitutions at Ser465/470 and Thr463/466 were poorly internalized, and Thr463 seemed to be the most important C-terminal Thr or Ser residue directly effecting receptor internalization. However, it is likely that other C3aR regions additionally participate in this negative feed-back mechanism since even mutants with multiple Ala substitutions still internalized to a limited degree. Interestingly, in a mutant with a single exchange of Ser449 to Ala, the signal transduction assessed by a Ca(2+) assay and [(35)S]GTP gamma S-binding on HEK cells transiently co-transfected with G-alpha 16 or G-alpha O, respectively, was severely impaired, indicating that this residue of C3aR is involved in G protein coupling. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/1127670
- author
- Settmacher, Britta ; Rheinheimer, Claudia ; Hamacher, Henning ; Ames, Robert S ; Wise, Alan ; Jenkinson, Lesley ; Bock, Daniel ; Schaefer, Myriam ; Kohl, Jorg and Klos, Andreas
- publishing date
- 2003
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- Complement, Receptor, Internalization, Inflammation, Regulation
- in
- European Journal of Immunology
- volume
- 33
- issue
- 4
- pages
- 920 - 927
- publisher
- John Wiley & Sons Inc.
- external identifiers
-
- pmid:12672058
- scopus:0038068127
- pmid:12672058
- ISSN
- 1521-4141
- DOI
- 10.1002/eji.200323293
- language
- English
- LU publication?
- no
- id
- 76defca9-7ab9-4ae4-8da6-af72805abeb9 (old id 1127670)
- alternative location
- http://www.ncbi.nlm.nih.gov/pubmed/12672058
- http://onlinelibrary.wiley.com/doi/10.1002/eji.200323293/abstract;jsessionid=82F53A0AB2941B4D6AA2D40D552DE71F.f02t01
- date added to LUP
- 2016-04-01 12:19:43
- date last changed
- 2022-01-27 02:07:17
@article{76defca9-7ab9-4ae4-8da6-af72805abeb9, abstract = {{The anaphylatoxic peptide C3a is a pro-inflammatory mediator generated during complement activation, whose specific G protein coupled receptor is expressed on granulocytes, monocytes, mast cells, activated lymphocytes, and in the nervous tissue. We have generated RBL-2H3 cell clones stably expressing mutants of the human C3a-receptor (C3aR) with combined alanine (Ala) substitutions of ten C-terminal serine (Ser) or threonine (Thr) residues, which may represent putative phosphorylation sites to characterize their role in ligand-induced C3aR internalization and signaling. Ser475/479 and Thr480/481 as well as Ser449 seemed not to be involved in ligand-induced receptor internalization. Either directly or by a conformational change they even "inhibit" C3aR internalization. In contrast, mutants with Ala substitutions at Ser465/470 and Thr463/466 were poorly internalized, and Thr463 seemed to be the most important C-terminal Thr or Ser residue directly effecting receptor internalization. However, it is likely that other C3aR regions additionally participate in this negative feed-back mechanism since even mutants with multiple Ala substitutions still internalized to a limited degree. Interestingly, in a mutant with a single exchange of Ser449 to Ala, the signal transduction assessed by a Ca(2+) assay and [(35)S]GTP gamma S-binding on HEK cells transiently co-transfected with G-alpha 16 or G-alpha O, respectively, was severely impaired, indicating that this residue of C3aR is involved in G protein coupling.}}, author = {{Settmacher, Britta and Rheinheimer, Claudia and Hamacher, Henning and Ames, Robert S and Wise, Alan and Jenkinson, Lesley and Bock, Daniel and Schaefer, Myriam and Kohl, Jorg and Klos, Andreas}}, issn = {{1521-4141}}, keywords = {{Complement; Receptor; Internalization; Inflammation; Regulation}}, language = {{eng}}, number = {{4}}, pages = {{920--927}}, publisher = {{John Wiley & Sons Inc.}}, series = {{European Journal of Immunology}}, title = {{Structure-function studies of the C3a-receptor: C-terminal serine and threonine residues which influence receptor internalization and signaling}}, url = {{http://dx.doi.org/10.1002/eji.200323293}}, doi = {{10.1002/eji.200323293}}, volume = {{33}}, year = {{2003}}, }