Lancellotti, S, Rutella, S ORCID: https://orcid.org/0000-0003-1970-7375, De Filippis, V, Pozzi, N, Rocca, B and De Cristofaro, R, 2008. Fibrinogen-elongated Chain Inhibits Thrombin-induced Platelet Response, Hindering the Interaction with Different Receptors. Journal of Biological Chemistry, 283 (44), pp. 30193-30204. ISSN 0021-9258
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Abstract
The expression of the elongated fibrinogen γ chain, termed γ′, derives from alternative splicing of mRNA and causes an insertion sequence of 20 amino acids. This insertion domain interacts with the anion-binding exosite (ABE)-II of thrombin. This study investigated whether and how γ′ chain binding to ABE-II affects thrombin interaction with its platelet receptors, i.e. glycoprotein Ibα (GpIbα), protease-activated receptor (PAR) 1, and PAR4. Both synthetic γ′ peptide and fibrinogen fragment D*, containing the elongated γ′ chain, inhibited thrombin-induced platelet aggregation up to 70%, with IC50 values of 42 ± 3.5 and 0.47 ± 0.03 μm, respectively. Solid-phase binding and spectrofluorimetric assays showed that both fragment D* and the synthetic γ′ peptide specifically bind to thrombin ABE-II and competitively inhibit the thrombin binding to GpIbα with a mean Ki ≈ 0.5 and ≈35 μm, respectively. Both these γ′ chain-containing ligands allosterically inhibited thrombin cleavage of a synthetic PAR1 peptide, of native PAR1 molecules on intact platelets, and of the synthetic chromogenic peptide d-Phe-pipecolyl-Arg-p-nitroanilide. PAR4 cleavage was unaffected. In summary, fibrinogen γ′ chain binds with high affinity to thrombin and inhibits with combined mechanisms the platelet response to thrombin. Thus, its variations in vivo may affect the hemostatic balance in arterial circulation.
Item Type: | Journal article |
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Publication Title: | Journal of Biological Chemistry |
Creators: | Lancellotti, S., Rutella, S., De Filippis, V., Pozzi, N., Rocca, B. and De Cristofaro, R. |
Publisher: | American Society for Biochemistry and Molecular Biology |
Date: | 31 October 2008 |
Volume: | 283 |
Number: | 44 |
ISSN: | 0021-9258 |
Identifiers: | Number Type 10.1074/jbc.M803659200 DOI |
Divisions: | Schools > School of Science and Technology |
Record created by: | Linda Sullivan |
Date Added: | 13 Sep 2016 14:52 |
Last Modified: | 09 Jun 2017 14:05 |
URI: | https://irep.ntu.ac.uk/id/eprint/28509 |
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