PMCCPMCCPMCC

Search tips
Search criteria 

Advanced

 
Logo of jexpmedHomeEditorsContactInstructions for Authors
 
J Exp Med. 1988 November 1; 168(5): 1699–1717.
PMCID: PMC2189104

Identification of distinct C3b and C4b recognition sites in the human C3b/C4b receptor (CR1, CD35) by deletion mutagenesis

Abstract

Complementary DNA clones encoding the NH2-terminal region of human CR1 have been isolated and sequenced. The deduced complete amino acid sequence of the F allotype of human CR1 contains 2,039 residues, including a 41-residue signal peptide, an extracellular domain of 1,930 residues, a 25-amino acid transmembrane domain, and a 43-amino acid cytoplasmic region. The extracellular domain is composed exclusively of 30 short consensus repeats (SCRs), characteristic of the family of C3/C4-binding proteins. The 28 NH2-terminal SCRs are organized as four long homologous repeats (LHRs) of seven SCRs each. The newly sequenced LHR, LHR-A, is 61% identical to LHR-B in the NH2-terminal two SCRs and greater than 99% identical in the COOH-terminal five SCRs. Eight cDNA clones were spliced to form a single construct, piABCD, that contained the entire CR1 coding sequence downstream of a cytomegalovirus promoter. COS cells transfected with piABCD transiently expressed recombinant CR1 that comigrated with the F allotype of erythrocyte CR1 on SDS-PAGE and that mediated rosette formation with sheep erythrocytes bearing C4b and C3b. Recombinant CR1 also had factor I-cofactor activity for cleavage of C3(ma). Analyses of six deletion mutants expressed in COS cells indicated that the NH2-terminal two SCRs of LHR- A contained a site determining C4 specificity and the NH2-terminal two SCRs of LHR-B and -C each had a site determining C3 specificity. The presence of these three distinct sites in CR1 may enable the receptor to interact multivalently with C4b/C3b and C3b/C3b complexes generated during activation of the classical and alternative pathways.

Full Text

The Full Text of this article is available as a PDF (1.4M).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Ross GD, Medof ME. Membrane complement receptors specific for bound fragments of C3. Adv Immunol. 1985;37:217–267. [PubMed]
  • Van Dyne S, Holers VM, Lublin DM, Atkinson JP. The polymorphism of the C3b/C4b receptor in the normal population and in patients with systemic lupus erythematosus. Clin Exp Immunol. 1987 Jun;68(3):570–579. [PubMed]
  • Klickstein LB, Wong WW, Smith JA, Weis JH, Wilson JG, Fearon DT. Human C3b/C4b receptor (CR1). Demonstration of long homologous repeating domains that are composed of the short consensus repeats characteristics of C3/C4 binding proteins. J Exp Med. 1987 Apr 1;165(4):1095–1112. [PMC free article] [PubMed]
  • Kristensen T, D'Eustachio P, Ogata RT, Chung LP, Reid KB, Tack BF. The superfamily of C3b/C4b-binding proteins. Fed Proc. 1987 May 15;46(7):2463–2469. [PubMed]
  • Chirgwin JM, Przybyla AE, MacDonald RJ, Rutter WJ. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. [PubMed]
  • Aviv H, Leder P. Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1408–1412. [PubMed]
  • Wong WW, Klickstein LB, Smith JA, Weis JH, Fearon DT. Identification of a partial cDNA clone for the human receptor for complement fragments C3b/C4b. Proc Natl Acad Sci U S A. 1985 Nov;82(22):7711–7715. [PubMed]
  • Wong WW, Kennedy CA, Bonaccio ET, Wilson JG, Klickstein LB, Weis JH, Fearon DT. Analysis of multiple restriction fragment length polymorphisms of the gene for the human complement receptor type I. Duplication of genomic sequences occurs in association with a high molecular mass receptor allotype. J Exp Med. 1986 Nov 1;164(5):1531–1546. [PMC free article] [PubMed]
  • Sanger F, Nicklen S, Coulson AR. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. [PubMed]
  • Yanisch-Perron C, Vieira J, Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. [PubMed]
  • Henikoff S. Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene. 1984 Jun;28(3):351–359. [PubMed]
  • Chen EY, Seeburg PH. Supercoil sequencing: a fast and simple method for sequencing plasmid DNA. DNA. 1985 Apr;4(2):165–170. [PubMed]
  • Devereux J, Haeberli P, Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. [PMC free article] [PubMed]
  • Seed B. An LFA-3 cDNA encodes a phospholipid-linked membrane protein homologous to its receptor CD2. Nature. 329(6142):840–842. [PubMed]
  • Lopata MA, Cleveland DW, Sollner-Webb B. High level transient expression of a chloramphenicol acetyl transferase gene by DEAE-dextran mediated DNA transfection coupled with a dimethyl sulfoxide or glycerol shock treatment. Nucleic Acids Res. 1984 Jul 25;12(14):5707–5717. [PMC free article] [PubMed]
  • Changelian PS, Jack RM, Collins LA, Fearon DT. PMA induces the ligand-independent internalization of CR1 on human neutrophils. J Immunol. 1985 Mar;134(3):1851–1858. [PubMed]
  • Fraker PJ, Speck JC., Jr Protein and cell membrane iodinations with a sparingly soluble chloroamide, 1,3,4,6-tetrachloro-3a,6a-diphrenylglycoluril. Biochem Biophys Res Commun. 1978 Feb 28;80(4):849–857. [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • Lambris JD, Scheiner O, Schulz TF, Alsenz J, Dierich MP. Coupling of C3b to erythrocytes by disulfide bond formation: preparation of EC3b for hemolytic and complement receptor assays. J Immunol Methods. 1983 Dec 30;65(3):277–283. [PubMed]
  • Hammer CH, Wirtz GH, Renfer L, Gresham HD, Tack BF. Large scale isolation of functionally active components of the human complement system. J Biol Chem. 1981 Apr 25;256(8):3995–4006. [PubMed]
  • Fearon DT. Purification of C3b inactivator and demonstration of its two polypeptide chain structure. J Immunol. 1977 Oct;119(4):1248–1252. [PubMed]
  • von Heijne G. A new method for predicting signal sequence cleavage sites. Nucleic Acids Res. 1986 Jun 11;14(11):4683–4690. [PMC free article] [PubMed]
  • Kozak M. Point mutations define a sequence flanking the AUG initiator codon that modulates translation by eukaryotic ribosomes. Cell. 1986 Jan 31;44(2):283–292. [PubMed]
  • Wong WW, Jack RM, Smith JA, Kennedy CA, Fearon DT. Rapid purification of the human C3b/C4b receptor (CR1) by monoclonal antibody affinity chromatography. J Immunol Methods. 1985 Oct 10;82(2):303–313. [PubMed]
  • Holers VM, Seya T, Brown E, O'Shea JJ, Atkinson JP. Structural and functional studies on the human C3b/C4b receptor (CR1) purified by affinity chromatography using a monoclonal antibody. Complement. 1986;3(2):63–78. [PubMed]
  • Selden RF, Howie KB, Rowe ME, Goodman HM, Moore DD. Human growth hormone as a reporter gene in regulation studies employing transient gene expression. Mol Cell Biol. 1986 Sep;6(9):3173–3179. [PMC free article] [PubMed]
  • Fearon DT, Kaneko I, Thomson GG. Membrane distribution and adsorptive endocytosis by C3b receptors on human polymorphonuclear leukocytes. J Exp Med. 1981 Jun 1;153(6):1615–1628. [PMC free article] [PubMed]
  • Fujita T, Nussenzweig V. The role of C4-binding protein and beta 1H in proteolysis of C4b and C3b. J Exp Med. 1979 Aug 1;150(2):267–276. [PMC free article] [PubMed]
  • Pangburn MK, Schreiber RD, Müller-Eberhard HJ. Human complement C3b inactivator: isolation, characterization, and demonstration of an absolute requirement for the serum protein beta1H for cleavage of C3b and C4b in solution. J Exp Med. 1977 Jul 1;146(1):257–270. [PMC free article] [PubMed]
  • Arnaout MA, Dana N, Melamed J, Medicus R, Colten HR. Low ionic strength or chemical cross-linking of monomeric C3b increases its binding affinity to the human complement C3b receptor. Immunology. 1983 Feb;48(2):229–237. [PubMed]
  • Gaither TA, Magrath IT, Berger M, Hammer CH, Novikovs L, Santaella M, Frank MM. Complement receptor expression by neoplastic and normal human cells. J Immunol. 1983 Aug;131(2):899–905. [PubMed]
  • Takata Y, Kinoshita T, Kozono H, Takeda J, Tanaka E, Hong K, Inoue K. Covalent association of C3b with C4b within C5 convertase of the classical complement pathway. J Exp Med. 1987 Jun 1;165(6):1494–1507. [PMC free article] [PubMed]
  • Wilson JG, Ratnoff WD, Schur PH, Fearon DT. Decreased expression of the C3b/C4b receptor (CR1) and the C3d receptor (CR2) on B lymphocytes and of CR1 on neutrophils of patients with systemic lupus erythematosus. Arthritis Rheum. 1986 Jun;29(6):739–747. [PubMed]
  • Edberg JC, Wright E, Taylor RP. Quantitative analyses of the binding of soluble complement-fixing antibody/dsDNA immune complexes to CR1 on human red blood cells. J Immunol. 1987 Dec 1;139(11):3739–3747. [PubMed]
  • Iida K, Mornaghi R, Nussenzweig V. Complement receptor (CR1) deficiency in erythrocytes from patients with systemic lupus erythematosus. J Exp Med. 1982 May 1;155(5):1427–1438. [PMC free article] [PubMed]
  • Wilson JG, Wong WW, Schur PH, Fearon DT. Mode of inheritance of decreased C3b receptors on erythrocytes of patients with systemic lupus erythematosus. N Engl J Med. 1982 Oct 14;307(16):981–986. [PubMed]

Articles from The Journal of Experimental Medicine are provided here courtesy of The Rockefeller University Press