Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1999 Jul 1;341(Pt 1):61–69.

Identification of human complement Factor H as a ligand for L-selectin.

R Malhotra 1, M Ward 1, R B Sim 1, M I Bird 1
PMCID: PMC1220330  PMID: 10377245

Abstract

The selectin family of adhesion molecules (E-, P- and L-selectins) is involved in leukocyte recruitment to sites of inflammation and tissue damage. Recently it has been shown that L-selectin is involved not only in leukocyte tethering and rolling, but also plays an important role in leukocyte activation. For example, glycosylation-dependent cell-adhesion molecule 1 (GlyCAM-1), a known ligand for L-selectin, has been shown to enhance beta2-integrin function. GlyCAM-1 is a secreted protein and is present in mouse serum at a concentration of approx. 1.5 microg/ml. There is no obvious GlyCAM-1 homologue in man and, to date, L-selectin ligand(s) from human serum have not been characterized. Therefore we have used L-selectin affinity chromatography, followed by ion-exchange chromatography, to isolate specific ligand(s) for L-selectin. Using this procedure, we have isolated three major glycoproteins of apparent molecular masses 170 kDa, 70kDa and 50 kDa. The 170 kDa protein band was digested with trypsin and peptides were analysed by delayed extraction matrix-assisted laser desorption ionization MS and protein database searching. The 170 kDa protein was identified as the human complement protein Factor H. Human Factor H, isolated by a different method, was shown to bind specifically to L-selectin in the presence of CaCl2, and binding was inhibited by anti-L-selectin antibodies, fucoidan and lipopolysaccharide. Only a part of the purified Factor H preparation bound to immobilized L-selectin. The interaction of Factor H with leukocyte L-selectin was shown to induce the secretion of tumour necrosis factor-alpha (TNF-alpha). Pretreatment of Factor H with sialidase reduced both the binding of L-selectin to Factor H and the Factor H-induced L-selectin-mediated TNF-alpha secretion by leukocytes. Taken together, these results demonstrate that a post-translationally modified form of human plasma Factor H is a potential physiological ligand for L-selectin.

Full Text

The Full Text of this article is available as a PDF (197.7 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Avery V. M., Gordon D. L. Characterization of factor H binding to human polymorphonuclear leukocytes. J Immunol. 1993 Nov 15;151(10):5545–5553. [PubMed] [Google Scholar]
  2. Bonner R., Shushan B. The characterization of proteins and peptides by automated methods. Rapid Commun Mass Spectrom. 1995;9(11):1067–1076. doi: 10.1002/rcm.1290091119. [DOI] [PubMed] [Google Scholar]
  3. Brown R. S., Lennon J. J. Mass resolution improvement by incorporation of pulsed ion extraction in a matrix-assisted laser desorption/ionization linear time-of-flight mass spectrometer. Anal Chem. 1995 Jul 1;67(13):1998–2003. doi: 10.1021/ac00109a015. [DOI] [PubMed] [Google Scholar]
  4. Clemmensen I. Interaction of tetranectin with sulphated polysaccharides and trypan blue. Scand J Clin Lab Invest. 1989 Dec;49(8):719–725. doi: 10.3109/00365518909091550. [DOI] [PubMed] [Google Scholar]
  5. Clemmensen I., Petersen L. C., Kluft C. Purification and characterization of a novel, oligomeric, plasminogen kringle 4 binding protein from human plasma: tetranectin. Eur J Biochem. 1986 Apr 15;156(2):327–333. doi: 10.1111/j.1432-1033.1986.tb09586.x. [DOI] [PubMed] [Google Scholar]
  6. Erdei A., Sim R. B. Complement factor H-binding protein of Raji cells and tonsil B lymphocytes. Biochem J. 1987 Aug 15;246(1):149–156. doi: 10.1042/bj2460149. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Giblin P. A., Hwang S. T., Katsumoto T. R., Rosen S. D. Ligation of L-selectin on T lymphocytes activates beta1 integrins and promotes adhesion to fibronectin. J Immunol. 1997 Oct 1;159(7):3498–3507. [PubMed] [Google Scholar]
  8. Harrison R. A., Lachmann P. J. Novel cleavage products of the third component of human complement. Mol Immunol. 1980 Feb;17(2):219–228. doi: 10.1016/0161-5890(80)90074-7. [DOI] [PubMed] [Google Scholar]
  9. Hellwage J., Kühn S., Zipfel P. F. The human complement regulatory factor-H-like protein 1, which represents a truncated form of factor H, displays cell-attachment activity. Biochem J. 1997 Sep 1;326(Pt 2):321–327. doi: 10.1042/bj3260321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hemmerich S., Leffler H., Rosen S. D. Structure of the O-glycans in GlyCAM-1, an endothelial-derived ligand for L-selectin. J Biol Chem. 1995 May 19;270(20):12035–12047. doi: 10.1074/jbc.270.20.12035. [DOI] [PubMed] [Google Scholar]
  11. Hwang S. T., Singer M. S., Giblin P. A., Yednock T. A., Bacon K. B., Simon S. I., Rosen S. D. GlyCAM-1, a physiologic ligand for L-selectin, activates beta 2 integrins on naive peripheral lymphocytes. J Exp Med. 1996 Oct 1;184(4):1343–1348. doi: 10.1084/jem.184.4.1343. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Iferroudjene D., Schouft M. T., Lemercier C., Gilbert D., Fontaine M. Evidence for an active hydrophobic form of factor H that is able to induce secretion of interleukin 1-beta or by human monocytes. Eur J Immunol. 1991 Apr;21(4):967–972. doi: 10.1002/eji.1830210416. [DOI] [PubMed] [Google Scholar]
  13. Jensen O. N., Podtelejnikov A. V., Mann M. Identification of the components of simple protein mixtures by high-accuracy peptide mass mapping and database searching. Anal Chem. 1997 Dec 1;69(23):4741–4750. doi: 10.1021/ac970896z. [DOI] [PubMed] [Google Scholar]
  14. Kansas G. S. Selectins and their ligands: current concepts and controversies. Blood. 1996 Nov 1;88(9):3259–3287. [PubMed] [Google Scholar]
  15. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  16. Lambris J., Papamichail M. A method for analysing lymphocyte surface antigens. J Immunol Methods. 1980;34(4):287–293. doi: 10.1016/0022-1759(80)90101-5. [DOI] [PubMed] [Google Scholar]
  17. Ley K. Gene-targeted mice in leukocyte adhesion research. Microcirculation. 1995 Aug;2(2):141–150. doi: 10.3109/10739689509146762. [DOI] [PubMed] [Google Scholar]
  18. Li F., Wilkins P. P., Crawley S., Weinstein J., Cummings R. D., McEver R. P. Post-translational modifications of recombinant P-selectin glycoprotein ligand-1 required for binding to P- and E-selectin. J Biol Chem. 1996 Feb 9;271(6):3255–3264. [PubMed] [Google Scholar]
  19. Malhotra R., Bird M. I. L-Selectin--a signalling receptor for lipopolysaccharide. Chem Biol. 1997 Aug;4(8):543–547. doi: 10.1016/s1074-5521(97)90240-4. [DOI] [PubMed] [Google Scholar]
  20. Malhotra R., Priest R., Bird M. I. Role for L-selectin in lipopolysaccharide-induced activation of neutrophils. Biochem J. 1996 Dec 1;320(Pt 2):589–593. doi: 10.1042/bj3200589. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Malhotra R., Taylor N. R., Bird M. I. Anionic phospholipids bind to L-selectin (but not E-selectin) at a site distinct from the carbohydrate-binding site. Biochem J. 1996 Feb 15;314(Pt 1):297–303. doi: 10.1042/bj3140297. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Mann M., Højrup P., Roepstorff P. Use of mass spectrometric molecular weight information to identify proteins in sequence databases. Biol Mass Spectrom. 1993 Jun;22(6):338–345. doi: 10.1002/bms.1200220605. [DOI] [PubMed] [Google Scholar]
  23. Mann M., Wilm M. Electrospray mass spectrometry for protein characterization. Trends Biochem Sci. 1995 Jun;20(6):219–224. doi: 10.1016/s0968-0004(00)89019-2. [DOI] [PubMed] [Google Scholar]
  24. McEver R. P., Cummings R. D. Perspectives series: cell adhesion in vascular biology. Role of PSGL-1 binding to selectins in leukocyte recruitment. J Clin Invest. 1997 Aug 1;100(3):485–491. doi: 10.1172/JCI119556. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. McEver R. P. Regulation of function and expression of P-selectin. Agents Actions Suppl. 1995;47:117–119. doi: 10.1007/978-3-0348-7343-7_10. [DOI] [PubMed] [Google Scholar]
  26. Nabil K., Rihn B., Jaurand M. C., Vignaud J. M., Ripoche J., Martinet Y., Martinet N. Identification of human complement factor H as a chemotactic protein for monocytes. Biochem J. 1997 Sep 1;326(Pt 2):377–383. doi: 10.1042/bj3260377. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Reid K. B. Proteins involved in the activation and control of the two pathways of human complement. Biochem Soc Trans. 1983 Jan;11(1):1–12. doi: 10.1042/bst0110001. [DOI] [PubMed] [Google Scholar]
  28. Ripoche J., Al Salihi A., Rousseaux J., Fontaine M. Isolation of two molecular populations of human complement factor H by hydrophobic affinity chromatography. Biochem J. 1984 Jul 1;221(1):89–96. doi: 10.1042/bj2210089. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Ripoche J., Day A. J., Harris T. J., Sim R. B. The complete amino acid sequence of human complement factor H. Biochem J. 1988 Jan 15;249(2):593–602. doi: 10.1042/bj2490593. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Ripoche J., Erdei A., Gilbert D., Al Salihi A., Sim R. B., Fontaine M. Two populations of complement factor H differ in their ability to bind to cell surfaces. Biochem J. 1988 Jul 15;253(2):475–480. doi: 10.1042/bj2530475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Rosen S. D., Bertozzi C. R. Two selectins converge on sulphate. Leukocyte adhesion. Curr Biol. 1996 Mar 1;6(3):261–264. doi: 10.1016/s0960-9822(02)00473-6. [DOI] [PubMed] [Google Scholar]
  32. Sim E., Palmer M. S., Puklavec M., Sim R. B. Monoclonal antibodies against the complement control protein factor H (beta 1 H). Biosci Rep. 1983 Dec;3(12):1119–1131. doi: 10.1007/BF01120205. [DOI] [PubMed] [Google Scholar]
  33. Sim E., Wood A. B., Hsiung L. M., Sim R. B. Pattern of degradation of human complement fragment, C3b. FEBS Lett. 1981 Sep 14;132(1):55–60. doi: 10.1016/0014-5793(81)80426-7. [DOI] [PubMed] [Google Scholar]
  34. Sim R. B., Day A. J., Moffatt B. E., Fontaine M. Complement factor I and cofactors in control of complement system convertase enzymes. Methods Enzymol. 1993;223:13–35. doi: 10.1016/0076-6879(93)23035-l. [DOI] [PubMed] [Google Scholar]
  35. Sim R. B., DiScipio R. G. Purification and structural studies on the complement-system control protein beta 1H (Factor H). Biochem J. 1982 Aug 1;205(2):285–293. doi: 10.1042/bj2050285. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Sim R. B., Moestrup S. K., Stuart G. R., Lynch N. J., Lu J., Schwaeble W. J., Malhotra R. Interaction of C1q and the collectins with the potential receptors calreticulin (cC1qR/collectin receptor) and megalin. Immunobiology. 1998 Aug;199(2):208–224. doi: 10.1016/s0171-2985(98)80028-4. [DOI] [PubMed] [Google Scholar]
  37. Wilm M., Mann M. Analytical properties of the nanoelectrospray ion source. Anal Chem. 1996 Jan 1;68(1):1–8. doi: 10.1021/ac9509519. [DOI] [PubMed] [Google Scholar]
  38. Wilm M., Shevchenko A., Houthaeve T., Breit S., Schweigerer L., Fotsis T., Mann M. Femtomole sequencing of proteins from polyacrylamide gels by nano-electrospray mass spectrometry. Nature. 1996 Feb 1;379(6564):466–469. doi: 10.1038/379466a0. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES