Effect of Sulfotyrosine and Negatively Charged Amino Acid of Leech-Derived Peptides on Binding and Inhibitory Activity Against Thrombin
Journal
Chembiochem : a European journal of chemical biology
Journal Volume
25
Journal Issue
3
Pages
e202300744
Date Issued
2024-02-01
Author(s)
Chen, Tzu-Yin
Shyur, Eileen
Ma, Tzu-Hsuan
Wijeyewickrema, Lakshmi
Lin, Sheng-Wei
Kao, Mu-Rong
Shie, Jiun-Jie
Chuang, Er-Yuan
Liou, Jing-Ping
Hsieh, Yves S Y
Abstract
Hirudins, natural sulfo(glyco)proteins, are clinical anticoagulants that directly inhibit thrombin, a key coagulation factor. Their potent thrombin inhibition primarily results from antagonistic interactions with both the catalytic and non-catalytic sites of thrombin. Hirudins often feature sulfate moieties on Tyr residues in their anionic C-terminus region, enabling strong interactions with thrombin exosite-I and effectively blocking its engagement with fibrinogen. Although sulfotyrosines have been identified in various hirudin variants, the precise relationship between sulfotyrosine and the number of negatively charged amino acids within the anionic-rich C-terminus peptide domain for the binding of thrombin has remained elusive. By using Fmoc-SPPS, hirudin dodecapeptides homologous to the C-terminus of hirudin variants from various leech species were successfully synthesized, and the effect of sulfotyrosine and the number of negatively charged amino acids on hirudin-thrombin interactions was investigated. Our findings did not reveal any synergistic effect between an increasing number of sulfotyrosines or negatively charged amino acids and their inhibitory activity on thrombin or fibrinolysis in the assays, despite a higher binding level toward thrombin in the sulfated dodecapeptide Hnip_Hirudin was observed in SPR analysis.
Subjects
hirudin; negatively charged amino acid; sulfotyrosine; thrombin
SDGs
Type
journal article