Skip to main content
PMC home page
Protein Science : A Publication of the Protein Society logoLink to Protein Science : A Publication of the Protein Society
. 1999 Dec;8(12):2663–2671. doi: 10.1110/ps.8.12.2663

Structure of the fibrinogen gamma-chain integrin binding and factor XIIIa cross-linking sites obtained through carrier protein driven crystallization.

S Ware 1, J P Donahue 1, J Hawiger 1, W F Anderson 1
PMCID: PMC2144222  PMID: 10631982

Abstract

The human fibrinogen gamma-chain C-terminal segment functions as the platelet integrin binding site as well as the Factor XIIIa cross-linking substrate and thus plays an important role in blood clot formation and stabilization. The three-dimensional structure of this segment has been determined using carrier protein driven crystallization. The C-terminal segment, gamma-(398-411), was attached to a linker sequence at the C-terminus of glutathione S-transferase and the structure of this fusion protein determined at 1.8 A resolution. Functional studies of the chimeric protein demonstrate that the fibrinogen sequence in the presence of the carrier protein retains its specific functions as ligand for platelet integrin alpha(IIb)beta3 (gpIIb/IIIa) and as a cross-linking substrate for Factor XIIIa. The structure obtained for the fibrinogen gamma-chain segment is not affected by crystal packing and can provide the missing links to the recently reported model of cross-linked fibrin.

Full Text

The Full Text of this article is available as a PDF (3.0 MB).

Selected References

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

  1. Bernstein F. C., Koetzle T. F., Williams G. J., Meyer E. F., Jr, Brice M. D., Rodgers J. R., Kennard O., Shimanouchi T., Tasumi M. The Protein Data Bank: a computer-based archival file for macromolecular structures. J Mol Biol. 1977 May 25;112(3):535–542. doi: 10.1016/s0022-2836(77)80200-3. [DOI] [PubMed] [Google Scholar]
  2. Blumenstein M., Matsueda G. R., Timmons S., Hawiger J. A beta-turn is present in the 392-411 segment of the human fibrinogen gamma-chain. Effects of structural changes in this segment on affinity to antibody 4A5. Biochemistry. 1992 Nov 10;31(44):10692–10698. doi: 10.1021/bi00159a008. [DOI] [PubMed] [Google Scholar]
  3. Brünger A. T., Krukowski A., Erickson J. W. Slow-cooling protocols for crystallographic refinement by simulated annealing. Acta Crystallogr A. 1990 Jul 1;46(Pt 7):585–593. doi: 10.1107/s0108767390002355. [DOI] [PubMed] [Google Scholar]
  4. Brünger A. T., Kuriyan J., Karplus M. Crystallographic R factor refinement by molecular dynamics. Science. 1987 Jan 23;235(4787):458–460. doi: 10.1126/science.235.4787.458. [DOI] [PubMed] [Google Scholar]
  5. Chen R., Doolittle R. F. - cross-linking sites in human and bovine fibrin. Biochemistry. 1971 Nov 23;10(24):4487–4491. doi: 10.1021/bi00800a021. [DOI] [PubMed] [Google Scholar]
  6. Chung D. W., Chan W. Y., Davie E. W. Characterization of a complementary deoxyribonucleic acid coding for the gamma chain of human fibrinogen. Biochemistry. 1983 Jun 21;22(13):3250–3256. doi: 10.1021/bi00282a033. [DOI] [PubMed] [Google Scholar]
  7. Donahue J. P., Patel H., Anderson W. F., Hawiger J. Three-dimensional structure of the platelet integrin recognition segment of the fibrinogen gamma chain obtained by carrier protein-driven crystallization. Proc Natl Acad Sci U S A. 1994 Dec 6;91(25):12178–12182. doi: 10.1073/pnas.91.25.12178. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Doolittle R. F., Everse S. J., Spraggon G. Human fibrinogen: anticipating a 3-dimensional structure. FASEB J. 1996 Nov;10(13):1464–1470. doi: 10.1096/fasebj.10.13.8940292. [DOI] [PubMed] [Google Scholar]
  9. Doolittle R. F. Structural aspects of the fibrinogen to fibrin conversion. Adv Protein Chem. 1973;27:1–109. doi: 10.1016/s0065-3233(08)60446-5. [DOI] [PubMed] [Google Scholar]
  10. Farrell D. H., Thiagarajan P., Chung D. W., Davie E. W. Role of fibrinogen alpha and gamma chain sites in platelet aggregation. Proc Natl Acad Sci U S A. 1992 Nov 15;89(22):10729–10732. doi: 10.1073/pnas.89.22.10729. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hawiger J. Adhesive ends of fibrinogen and its antiadhesive peptides: the end of a saga? Semin Hematol. 1995 Apr;32(2):99–109. [PubMed] [Google Scholar]
  12. Jiang J. S., Brünger A. T. Protein hydration observed by X-ray diffraction. Solvation properties of penicillopepsin and neuraminidase crystal structures. J Mol Biol. 1994 Oct 14;243(1):100–115. doi: 10.1006/jmbi.1994.1633. [DOI] [PubMed] [Google Scholar]
  13. Kleywegt G. J., Brünger A. T. Checking your imagination: applications of the free R value. Structure. 1996 Aug 15;4(8):897–904. doi: 10.1016/s0969-2126(96)00097-4. [DOI] [PubMed] [Google Scholar]
  14. Kloczewiak M., Timmons S., Bednarek M. A., Sakon M., Hawiger J. Platelet receptor recognition domain on the gamma chain of human fibrinogen and its synthetic peptide analogues. Biochemistry. 1989 Apr 4;28(7):2915–2919. doi: 10.1021/bi00433a025. [DOI] [PubMed] [Google Scholar]
  15. Kloczewiak M., Timmons S., Lukas T. J., Hawiger J. Platelet receptor recognition site on human fibrinogen. Synthesis and structure-function relationship of peptides corresponding to the carboxy-terminal segment of the gamma chain. Biochemistry. 1984 Apr 10;23(8):1767–1774. doi: 10.1021/bi00303a028. [DOI] [PubMed] [Google Scholar]
  16. Lim K., Ho J. X., Keeling K., Gilliland G. L., Ji X., Rüker F., Carter D. C. Three-dimensional structure of Schistosoma japonicum glutathione S-transferase fused with a six-amino acid conserved neutralizing epitope of gp41 from HIV. Protein Sci. 1994 Dec;3(12):2233–2244. doi: 10.1002/pro.5560031209. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Mayo K. H., Fan F. Integrin receptor GPIIb/IIIa bound state conformation of the fibrinogen gamma-chain C-terminal peptide 400-411: NMR and transfer NOE studies. Biochemistry. 1996 Apr 9;35(14):4434–4444. doi: 10.1021/bi952485n. [DOI] [PubMed] [Google Scholar]
  18. McTigue M. A., Williams D. R., Tainer J. A. Crystal structures of a schistosomal drug and vaccine target: glutathione S-transferase from Schistosoma japonica and its complex with the leading antischistosomal drug praziquantel. J Mol Biol. 1995 Feb 10;246(1):21–27. doi: 10.1006/jmbi.1994.0061. [DOI] [PubMed] [Google Scholar]
  19. Merritt E. A., Murphy M. E. Raster3D Version 2.0. A program for photorealistic molecular graphics. Acta Crystallogr D Biol Crystallogr. 1994 Nov 1;50(Pt 6):869–873. doi: 10.1107/S0907444994006396. [DOI] [PubMed] [Google Scholar]
  20. Peerschke E. I., Francis C. W., Marder V. J. Fibrinogen binding to human blood platelets: effect of gamma chain carboxyterminal structure and length. Blood. 1986 Feb;67(2):385–390. [PubMed] [Google Scholar]
  21. Phillips D. R., Fitzgerald L., Parise L., Steiner B. Platelet membrane glycoprotein IIb-IIIa complex: purification, characterization, and reconstitution into phospholipid vesicles. Methods Enzymol. 1992;215:244–263. doi: 10.1016/0076-6879(92)15068-n. [DOI] [PubMed] [Google Scholar]
  22. Purves L., Purves M., Brandt W. Cleavage of fibrin-derived D-dimer into monomers by endopeptidase from puff adder venom (Bitis arietans) acting at cross-linked sites of the gamma-chain. Sequence of carboxy-terminal cyanogen bromide gamma-chain fragments. Biochemistry. 1987 Jul 28;26(15):4640–4646. doi: 10.1021/bi00389a008. [DOI] [PubMed] [Google Scholar]
  23. Rice L. M., Brünger A. T. Torsion angle dynamics: reduced variable conformational sampling enhances crystallographic structure refinement. Proteins. 1994 Aug;19(4):277–290. doi: 10.1002/prot.340190403. [DOI] [PubMed] [Google Scholar]
  24. Rossjohn J., McKinstry W. J., Oakley A. J., Verger D., Flanagan J., Chelvanayagam G., Tan K. L., Board P. G., Parker M. W. Human theta class glutathione transferase: the crystal structure reveals a sulfate-binding pocket within a buried active site. Structure. 1998 Mar 15;6(3):309–322. doi: 10.1016/s0969-2126(98)00034-3. [DOI] [PubMed] [Google Scholar]
  25. Rossmann M. G. The molecular replacement method. Acta Crystallogr A. 1990 Feb 1;46(Pt 2):73–82. doi: 10.1107/s0108767389009815. [DOI] [PubMed] [Google Scholar]
  26. Smith D. B., Johnson K. S. Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. Gene. 1988 Jul 15;67(1):31–40. doi: 10.1016/0378-1119(88)90005-4. [DOI] [PubMed] [Google Scholar]
  27. Spraggon G., Everse S. J., Doolittle R. F. Crystal structures of fragment D from human fibrinogen and its crosslinked counterpart from fibrin. Nature. 1997 Oct 2;389(6650):455–462. doi: 10.1038/38947. [DOI] [PubMed] [Google Scholar]
  28. Strong D. D., Laudano A. P., Hawiger J., Doolittle R. F. Isolation, characterization, and synthesis of peptides from human fibrinogen that block the staphylococcal clumping reaction and construction of a synthetic clumping particle. Biochemistry. 1982 Mar 16;21(6):1414–1420. doi: 10.1021/bi00535a048. [DOI] [PubMed] [Google Scholar]
  29. Yee V. C., Pratt K. P., Côté H. C., Trong I. L., Chung D. W., Davie E. W., Stenkamp R. E., Teller D. C. Crystal structure of a 30 kDa C-terminal fragment from the gamma chain of human fibrinogen. Structure. 1997 Jan 15;5(1):125–138. doi: 10.1016/s0969-2126(97)00171-8. [DOI] [PubMed] [Google Scholar]

Articles from Protein Science : A Publication of the Protein Society are provided here courtesy of The Protein Society

RESOURCES