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The Journal of experimental medicine
Feb 05, 2001;193 (3): 281-95.

Binding of C4b-binding protein to porin: a molecular mechanism of serum resistance of Neisseria gonorrhoeae.

S Ram, M Cullinane, A M Blom, S Gulati, D P McQuillen, B G Monks, C O'Connell, R Boden, C Elkins, M K Pangburn, B Dahlbäck, P A Rice
Author Information
  1. S Ram: Evans Biomedical Research Center, Boston Medical Center, Boston, Massachusetts 02118, USA. sram@bu.edu
PMID: 11157049 DOI: 10.1084/jem.193.3.281

Abstract

We screened 29 strains of Neisseria gonorrhoeae and found 16/21 strains that resisted killing by normal human serum and 0/8 serum sensitive strains that bound the complement regulator, C4b-binding protein (C4bp). Microbial surface-bound C4bp demonstrated cofactor activity. We constructed gonococcal strains with hybrid porin (Por) molecules derived from each of the major serogroups (Por1A and Por1B) of N. gonorrhoeae, and showed that the loop 1 of Por1A is required for C4bp binding. Por1B loops 5 and 7 of serum-resistant gonococci together formed a negatively charged C4bp-binding domain. C4bp-Por1B interactions were ionic in nature (inhibited by high salt or by heparin), whereas the C4bp-Por1A bond was hydrophobic. Only recombinant C4bp mutant molecules containing the NH2-terminal alpha-chain short consensus repeat (SCR1) bound to both Por1A and Por1B gonococci, suggesting that SCR1 contained Por binding sites. C4bp alpha-chain monomers did not bind gonococci, indicating that the polymeric form of C4bp was required for binding. Using fAb fragments against C4bp SCR1, C4bp binding to Por1A and Por1B strains was inhibited in a complement-dependent serum bactericidal assay. This resulted in complete killing of these otherwise fully serum resistant strains in only 10% normal serum, underscoring the importance of C4bp in mediating gonococcal serum resistance.

References

  1. J Exp Med. 1979 Aug 1;150(2):267-76 [PMID: 458376]
  2. Br J Vener Dis. 1979 Aug;55(4):265-70 [PMID: 114196]
  3. J Gen Microbiol. 1979 Sep;114(1):169-77 [PMID: 42666]
  4. Proc Natl Acad Sci U S A. 1979 Dec;76(12):6596-600 [PMID: 293746]
  5. J Immunol. 1980 May;124(5):2105-9 [PMID: 6767783]
  6. J Gen Microbiol. 1981 Feb;122(2):235-45 [PMID: 6798160]
  7. J Gen Microbiol. 1981 Nov;127(1):213-7 [PMID: 6802934]
  8. J Gen Microbiol. 1982 Jan;128(1):13-22 [PMID: 6806436]
  9. Cell. 1982 Aug;30(1):45-52 [PMID: 6127165]
  10. FEMS Microbiol Lett. 1992 Jul 15;73(3):283-9 [PMID: 1330818]
  11. Mol Microbiol. 1992 Sep;6(18):2617-28 [PMID: 1280317]
  12. Infect Immun. 1993 Jun;61(6):2273-6 [PMID: 8500868]
  13. Gene. 1994 Jan 28;138(1-2):43-50 [PMID: 8125316]
  14. Immunol Today. 1994 Mar;15(3):121-6 [PMID: 8172644]
  15. Methods Enzymol. 1994;236:137-47 [PMID: 7968606]
  16. J Biol Chem. 1994 Dec 2;269(48):30345-51 [PMID: 7982947]
  17. J Immunol. 1995 Jan 1;154(1):375-86 [PMID: 7995956]
  18. Infect Immun. 1983 May;40(2):816-9 [PMID: 6404835]
  19. Proc Natl Acad Sci U S A. 1983 Jun;80(11):3461-5 [PMID: 6222381]
  20. Biochem J. 1983 Mar 1;209(3):847-56 [PMID: 6223625]
  21. Medicine (Baltimore). 1983 Nov;62(6):395-406 [PMID: 6415361]
  22. FEBS Lett. 1983 Nov 28;164(1):135-8 [PMID: 6653778]
  23. J Infect Dis. 1984 Jul;150(1):44-8 [PMID: 6431013]
  24. J Biol Chem. 1984 Oct 10;259(19):11631-4 [PMID: 6480575]
  25. J Infect Dis. 1985 Aug;152(2):339-43 [PMID: 3928769]
  26. J Infect Dis. 1985 Nov;152(5):990-1001 [PMID: 2413148]
  27. Infect Immun. 1985 Dec;50(3):672-7 [PMID: 3934078]
  28. J Infect Dis. 1986 Mar;153(3):520-6 [PMID: 3081657]
  29. Genitourin Med. 1986 Jun;62(3):145-50 [PMID: 3089903]
  30. J Clin Invest. 1987 Jul;80(1):78-87 [PMID: 2439546]
  31. Infect Immun. 1988 Feb;56(2):499-504 [PMID: 3123395]
  32. Am J Epidemiol. 1999 Feb 15;149(4):353-8 [PMID: 10025478]
  33. J Biol Chem. 1999 Jul 2;274(27):19237-45 [PMID: 10383431]
  34. Immunopharmacology. 1999 May;42(1-3):53-60 [PMID: 10408366]
  35. Hum Reprod. 1988 Apr;3(3):289-300 [PMID: 3286677]
  36. J Biol Chem. 1988 Sep 5;263(25):12759-64 [PMID: 2970465]
  37. Proc Natl Acad Sci U S A. 1988 Sep;85(18):6841-5 [PMID: 2457919]
  38. J Exp Med. 1988 Sep 1;168(3):949-69 [PMID: 2844953]
  39. J Exp Med. 1988 Nov 1;168(5):1883-97 [PMID: 3141555]
  40. Microb Pathog. 1988 Oct;5(4):303-9 [PMID: 3148816]
  41. J Clin Invest. 1989 May;83(5):1472-6 [PMID: 2496142]
  42. Clin Microbiol Rev. 1989 Apr;2 Suppl:S92-103 [PMID: 2497967]
  43. J Exp Med. 1989 Jun 1;169(6):2199-209 [PMID: 2499656]
  44. Gene. 1989 Apr 15;77(1):51-9 [PMID: 2744487]
  45. J Gen Microbiol. 1988 Dec;134(12):3295-306 [PMID: 3151997]
  46. Biochem Biophys Res Commun. 1989 Nov 30;165(1):138-44 [PMID: 2590215]
  47. J Infect Dis. 1990 Aug;162(2):506-12 [PMID: 1695655]
  48. Mol Microbiol. 1990 Jun;4(6):1009-18 [PMID: 2170812]
  49. Thromb Haemost. 1990 Oct 22;64(2):245-50 [PMID: 2148654]
  50. Infect Immun. 1991 Apr;59(4):1278-84 [PMID: 1706317]
  51. Eur J Epidemiol. 1991 Jul;7(4):311-9 [PMID: 1717310]
  52. Lancet. 1991 Nov 16;338(8777):1272 [PMID: 1682666]
  53. Genitourin Med. 1992 Feb;68(1):16-9 [PMID: 1548006]
  54. Hum Reprod. 1994 Sep;9(9):1652-9 [PMID: 7836514]
  55. Mol Biol Evol. 1995 May;12(3):363-70 [PMID: 7739379]
  56. Br J Biomed Sci. 1995 Jun;52(2):87-92 [PMID: 8520255]
  57. J Clin Endocrinol Metab. 1996 Apr;81(4):1641-9 [PMID: 8636381]
  58. J Immunol. 1996 Oct 1;157(7):3021-9 [PMID: 8816411]
  59. J Infect Dis. 1996 Oct;174(4):760-7 [PMID: 8843214]
  60. Biochem J. 1997 Apr 15;323 ( Pt 2):469-75 [PMID: 9163340]
  61. J Bacteriol. 1997 Jun;179(11):3541-8 [PMID: 9171398]
  62. Infect Immun. 1997 Sep;65(9):3638-43 [PMID: 9284130]
  63. J Exp Med. 1998 Mar 2;187(5):743-52 [PMID: 9480984]
  64. Ann N Y Acad Sci. 1997 Dec 29;830:353-60 [PMID: 9616695]
  65. Proteins. 1998 Jun 1;31(4):391-405 [PMID: 9626699]
  66. J Exp Med. 1998 Aug 17;188(4):671-80 [PMID: 9705949]
  67. J Immunol. 1998 Oct 1;161(7):3711-8 [PMID: 9759896]
  68. J Immunol. 1998 Nov 1;161(9):4894-901 [PMID: 9794423]
  69. J Infect Dis. 1999 Jan;179(1):124-35 [PMID: 9841831]
  70. J Infect Dis. 1999 Feb;179(2):371-81 [PMID: 9878021]
  71. J Bacteriol. 1999 Sep;181(18):5591-9 [PMID: 10482498]
  72. Br J Vener Dis. 1961 Jun;37:145-57 [PMID: 13741074]
  73. Eur J Immunol. 2000 Apr;30(4):1243-53 [PMID: 10760814]
  74. Pediatr Infect Dis J. 2000 Apr;19(4):346-9 [PMID: 10783027]
  75. J Immunol. 2000 May 15;164(10):5328-36 [PMID: 10799895]
  76. Nature. 1970 Jul 25;227(5256):382-4 [PMID: 4987874]
  77. J Infect Dis. 1976 Feb;133(2):113-25 [PMID: 812926]
  78. J Exp Med. 1978 Jul 1;148(1):207-22 [PMID: 670886]
  79. J Lab Clin Med. 1978 Aug;92(2):211-20 [PMID: 98602]
  80. J Exp Med. 1978 Oct 1;148(4):1044-51 [PMID: 702059]
  81. Fertil Steril. 1979 Jul;32(1):61-6 [PMID: 456632]

Grants

  1. R37 AI032725/NIAID NIH HHS
  2. R37 DK035081/NIDDK NIH HHS
  3. R01 DK035081/NIDDK NIH HHS
  4. R56 AI032725/NIAID NIH HHS
  5. DK35081/NIDDK NIH HHS
  6. R01 AI032725/NIAID NIH HHS
  7. AI32725/NIAID NIH HHS

MeSH Term

Amino Acid Sequence
Base Sequence
Cell Line
Complement C4
Complement C4b
Complement Inactivator Proteins
Glycoproteins
Humans
Molecular Sequence Data
Neisseria gonorrhoeae
Peptide Fragments
Porins
Protein S
Receptors, Complement

Chemicals

Complement C4
Complement Inactivator Proteins
Glycoproteins
Peptide Fragments
Porins
Protein S
Receptors, Complement
complement C4c
porin protein, Neisseria
Complement C4b
complement C4d
Journal Article Research Support, U.S. Gov't, P.H.S.
Article has an altmetric score of 9

Word Cloud

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