Studies on Anticoagulation Factor Ⅱ from Agkistrodon Acutus Venom by Rare Earth Ion Fluorescence Probes

Abstract

Abstract: The anticoagulation factor Ⅱ (ACF-Ⅱ) from Agkistrodon acutus venom has two Ca ²⁺ binding sites and Ca ²⁺ ions enhance the intrinsic fluorescence of ACF-Ⅱ. Rare earth ions (Nd ³⁺, Sm³⁺, Eu ³⁺, Gd ³⁺, Tb ³⁺ ) can replace the Ca ²⁺ ions in ACF-Ⅱ and have significant quenching effects on the fluorescence of ACF-Ⅱ. The characteristic fluorescence of Tb ³⁺ is enhanced through the nonradiative energy transfer from Trp residues in ACF-Ⅱ to bound Tb ³⁺. The fluorescence titration of ACF-Ⅱ with rare earth ions demonstrates that ACF-Ⅱ has two RE ³⁺ binding sites, and the rare earth ions and Ca ²⁺ ions bind to ACF-Ⅱ competitively in the same two sites. Although the ionic radii of Nd ³⁺, Sm ³⁺, Eu ³⁺, Gd ³⁺ and Tb ³⁺ are different, their apparent association constant K₁ or K₂ are similar, respectively. Linear free energy relationships for Tb ³⁺ and RE ³⁺ (RE=Nd, Sm, Eu, Gd) indicate that there is not an obvious size restriction for the binding of lanthanide ions at the two binding sites of ACF-Ⅱ, which reveals the conformational flexibility of the two binding sites in ACF-Ⅱ. This conformational flexibility offers a possibility for Ca ²⁺ ions, when they take place in the inducing conformational changes of ACF-Ⅱ and the binding of ACF-Ⅱ with the activated coagulation factor Ⅹ.

Key words: Rare earth ions, Anticoagulation factor Ⅱ, Fluorescence probe, Metal ion binding site

CLC Number:

O657.3

TrendMD:

XU Xiao-Long, LIU Qing-Liang, XIE Yong-Shu . Studies on Anticoagulation Factor Ⅱ from Agkistrodon Acutus Venom by Rare Earth Ion Fluorescence Probes[J]. Chem. J. Chinese Universities, 2001, 22(11): 1807.

References

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