The effect of lanthanide and transition metal ions on the fluorescence spectra of the anticoagulation factor(ACF)of snake venom has been studied.It is postulated that the quenching of tryptophan residue fluorescence b...The effect of lanthanide and transition metal ions on the fluorescence spectra of the anticoagulation factor(ACF)of snake venom has been studied.It is postulated that the quenching of tryptophan residue fluorescence by lanthanide and transition metal ions is caused by the metal ion-induced conformation change of ACF.展开更多
It was observed that rare earth ions (Nd 3+, Sm 3+, Eu 3+, Gd 3+, Tb 3+) have significant quenching effects on the fluorescence of anticoagulation factor I (ACF I). The results of the fluorescence titra...It was observed that rare earth ions (Nd 3+, Sm 3+, Eu 3+, Gd 3+, Tb 3+) have significant quenching effects on the fluorescence of anticoagulation factor I (ACF I). The results of the fluorescence titration of ACF I with rare earth ions demonstrate that ACF I has two RE 3+-binding sites, and the rare earth ions and Ca 2+ bind to ACF I competitively in the two similar sites. The association constants K 1 and K 2 of ACF I with each rare earth ions (Nd 3+, Sm 3+, Eu 3+, Gd 3+, Tb 3+) are close to each other, which indicates the structural similarity of the two binding sites in ACF I. Although the ionic radii of Nd 3+, Sm 3+, Eu 3+, Gd 3+ and Tb 3+ are different, both their K 1 and K 2 are similar, respectively. This reveals the conformational flexibility of the two binding sites in ACF I, which offers a possibility for Ca 2+ to take play in the inducing conformational changes of ACF I and the promoting the binding of ACF I with activated coagulation factor X.展开更多
The effects of Pr3+, Gd3+ ions on the H-1 NMR spectrum of anticoagulation factor (ACF) from snake venom were investigated. It was observed that the quartet peaks at delta 4.16 and the triplet peaks at delta 1.37 are g...The effects of Pr3+, Gd3+ ions on the H-1 NMR spectrum of anticoagulation factor (ACF) from snake venom were investigated. It was observed that the quartet peaks at delta 4.16 and the triplet peaks at delta 1.37 are gradually broadened with the increase of Pr3+ ion content, but the broadening effects of Pr3+ ion on the two single peaks at delta 1.99 and delta 2.29 were not observed obviously, and the obvious chemical shifts of all peaks induced by Pr3+ were not found. Two peaks contributed to alphaH and PH of the Ala side chain in ACF are broadened by Gd3+ ion, while two single peaks at delta 2.29 and delta 2.29 are not affected basically by Gd3+ ion. This result proves that the distance between the Ca2+-binding site in ACF and aH or PH of the Ala side chain in ACF is shorter than that between the Ca2+-binding site in ACF and proton in Met residue.展开更多
Anticoagulation factor I (ACF I) from the venom of Agki^strodon acutus is a binding protein to activated coagulation factor X (FXa) and possesses marked anticoagulant activity. Single ACF I molecule has been succe...Anticoagulation factor I (ACF I) from the venom of Agki^strodon acutus is a binding protein to activated coagulation factor X (FXa) and possesses marked anticoagulant activity. Single ACF I molecule has been successfully imaged in air by tapping mode atomic force microscopy (AFM) with high resolution using glutaraldehyde as a coupling agent. The physical adsorption and covalent binding of ACF I onto the mica show very different surface topographies. The former exhibits the characteristic strand like structure with much less reproducibility, the latter displays a elliptic granular structure with better reproducibility, which suggests that the stability of ACF I molecules on the mica is enhanced by covalent bonding in the presence of glutaraldehyde. A small scale AFM amplitude mode image clearly shows that the covalently bonded ACF I molecule by glutaraldehyde has olive shape structure with an average size of 7 4 nm×3 6 nm×3 1 nm, which is very similar to the size determined from the crystal structure of ACF I.展开更多
基金Project supported by the National Natural Science Foundation of China
文摘The effect of lanthanide and transition metal ions on the fluorescence spectra of the anticoagulation factor(ACF)of snake venom has been studied.It is postulated that the quenching of tryptophan residue fluorescence by lanthanide and transition metal ions is caused by the metal ion-induced conformation change of ACF.
文摘It was observed that rare earth ions (Nd 3+, Sm 3+, Eu 3+, Gd 3+, Tb 3+) have significant quenching effects on the fluorescence of anticoagulation factor I (ACF I). The results of the fluorescence titration of ACF I with rare earth ions demonstrate that ACF I has two RE 3+-binding sites, and the rare earth ions and Ca 2+ bind to ACF I competitively in the two similar sites. The association constants K 1 and K 2 of ACF I with each rare earth ions (Nd 3+, Sm 3+, Eu 3+, Gd 3+, Tb 3+) are close to each other, which indicates the structural similarity of the two binding sites in ACF I. Although the ionic radii of Nd 3+, Sm 3+, Eu 3+, Gd 3+ and Tb 3+ are different, both their K 1 and K 2 are similar, respectively. This reveals the conformational flexibility of the two binding sites in ACF I, which offers a possibility for Ca 2+ to take play in the inducing conformational changes of ACF I and the promoting the binding of ACF I with activated coagulation factor X.
文摘The effects of Pr3+, Gd3+ ions on the H-1 NMR spectrum of anticoagulation factor (ACF) from snake venom were investigated. It was observed that the quartet peaks at delta 4.16 and the triplet peaks at delta 1.37 are gradually broadened with the increase of Pr3+ ion content, but the broadening effects of Pr3+ ion on the two single peaks at delta 1.99 and delta 2.29 were not observed obviously, and the obvious chemical shifts of all peaks induced by Pr3+ were not found. Two peaks contributed to alphaH and PH of the Ala side chain in ACF are broadened by Gd3+ ion, while two single peaks at delta 2.29 and delta 2.29 are not affected basically by Gd3+ ion. This result proves that the distance between the Ca2+-binding site in ACF and aH or PH of the Ala side chain in ACF is shorter than that between the Ca2+-binding site in ACF and proton in Met residue.
文摘Anticoagulation factor I (ACF I) from the venom of Agki^strodon acutus is a binding protein to activated coagulation factor X (FXa) and possesses marked anticoagulant activity. Single ACF I molecule has been successfully imaged in air by tapping mode atomic force microscopy (AFM) with high resolution using glutaraldehyde as a coupling agent. The physical adsorption and covalent binding of ACF I onto the mica show very different surface topographies. The former exhibits the characteristic strand like structure with much less reproducibility, the latter displays a elliptic granular structure with better reproducibility, which suggests that the stability of ACF I molecules on the mica is enhanced by covalent bonding in the presence of glutaraldehyde. A small scale AFM amplitude mode image clearly shows that the covalently bonded ACF I molecule by glutaraldehyde has olive shape structure with an average size of 7 4 nm×3 6 nm×3 1 nm, which is very similar to the size determined from the crystal structure of ACF I.
