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. 1997 Sep 1;16(17):5151–5161. doi: 10.1093/emboj/16.17.5151

X-ray structure of antistasin at 1.9 A resolution and its modelled complex with blood coagulation factor Xa.

R Lapatto 1, U Krengel 1, H A Schreuder 1, A Arkema 1, B de Boer 1, K H Kalk 1, W G Hol 1, P D Grootenhuis 1, J W Mulders 1, R Dijkema 1, H J Theunissen 1, B W Dijkstra 1
PMCID: PMC1170148  PMID: 9311976

Abstract

The three-dimensional structure of antistasin, a potent inhibitor of blood coagulation factor Xa, from the Mexican leech Haementeria officinalis was determined at 1.9 A resolution by X-ray crystallography. The structure reveals a novel protein fold composed of two homologous domains, each resembling the structure of hirustasin, a related 55-residue protease inhibitor. However, hirustasin has a different overall shape than the individual antistasin domains, it contains four rather than two beta-strands, and does not inhibit factor Xa. The two antistasin domains can be subdivided into two similarly sized subdomains with different relative orientations. Consequently, the domain shapes are different, the N-terminal domain being wedge-shaped and the C-terminal domain flat. Docking studies suggest that differences in domain shape enable the N-terminal, but not C-terminal, domain of antistasin to bind and inhibit factor Xa, even though both have a very similar reactive site. Furthermore, a putative exosite binding region could be defined in the N-terminal domain of antistasin, comprising residues 15-17, which is likely to interact with a cluster of positively charged residues on the factor Xa surface (Arg222/Lys223/Lys224). This exosite binding region explains the specificity and inhibitory potency of antistasin towards factor Xa. In the C-terminal domain of antistasin, these exosite interactions are prevented due to the different overall shape of this domain.

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Selected References

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