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 AJAC  Vol.4 No.12 , December 2013
Studies on the Binding Mechanism of VB1 and VB9 with Trypsin
Abstract: The binding characteristics of vitamin B1 (VB1) and vitamin B9 (VB9) with trypsin were investigated by fluorescence spectrometry and UV/vis spectrophotometry under simulated physiological conditions. With the addition of VB1 or VB9, the intrinsic fluorescence emission intensity of trypsin was quenched by the nonradiative energy transfer mechanism. The fluorescence quenching process of trypsin may be mainly governed by a static quenching mechanism. The binding parameters such as the binding constants and the number of binding sites can be evaluated by fluorescence quenching experiments. The numbers of the apparent binding constant Kb of VB1-trypsin at different temperatures were 0.4948 and 4.8340 × 104 L/mol and the numbers of binding sites n were 0.9359 and 1.1820. Similarly, the numbers of the apparent binding constant Kb of VB9-trypsin at different temperatures were 5.9310 and 13.040 × 104 L/mol and the numbers of binding sites n were 0.9908 and 1.0750. The thermodynamic parameters, with a negative value of ΔG, revealed that the bindings are spontaneous processes and the positive values for both enthalpy change (ΔH) and entropy change (ΔS) indicate that the binding powers of VB1 and VB9 with trypsin are mainly hydrophobic interactions. And synchronous spectrums were used to study the conformational change of trypsin. In addition, the binding distances of VB1-trypsin and VB9-trypsin were estimated to be 0.55 nm and 0.87 nm according to the Förster’s resonance energy transfer theory.
Cite this paper: Y. Gao, C. Shao, W. Ji, M. Xiao, F. Yi, T. Zhou and Y. Zi, "Studies on the Binding Mechanism of VB1 and VB9 with Trypsin," American Journal of Analytical Chemistry, Vol. 4 No. 12, 2013, pp. 771-775. doi: 10.4236/ajac.2013.412094.
References

[1]   H. B. Xue, C. Y. Li, L. C. Gao and G. Q. Zhang, “The Interaction of VB1 and Bovine Serum Albumin,” Chinese Journal of Analysis Laboratory, Vol. 30, No. 4, 2011, pp. 111-114.

[2]   H. J. Liu, P. Li, Y. D. Zhang, C. Guo, J. Y. Deng, J. W. Cai and B. L. Liu, “Spectroscopic Study on Binding of Folic Acid to Human Serum Albumin,” Spectroscopy and Spectral Analysis, Vol. 29, No. 7, 2009, pp. 1915-1919.

[3]   Y. H. Shang, H. Li, J. J. Sun and M. Y. Zhang, “Study on the Interaction of Bovine Serum Albumin with Riboflavin by Fluorescence Spectroscopy,” Journal of Anal Ytical Science, Vol. 26, No. 1, 2010, pp. 67-70.

[4]   L. Q. Sheng, X. Y. Yan, H. J. Xu, H. W. Tong and S. M. Liu, “Study on the Interaction between BSA and Nicotine,” Spectroscopy and Spectral Analysis, Vol. 27, No. 2, 2007, pp. 306-308.

[5]   A. M. Zhang, “Study on the Interaction Between VB2 and Serum Albumin by Fluorescence Spectroscopy,” Journal of Nanchang University, Vol. 30, Suppl., 2006, pp. 1087-1088.

[6]   Y. H. Fan, F. Feng, Z. Z. Chen and S. M. Shuang, “Studies on Interaction Between Bovine Serum Albumin and Vitamin B12 by Fluorescence Spectrometry,” Chinese Journal of Spectroscopy Laboratory, Vol. 28, No. 3, 2011, pp. 1331-1335.

[7]   H. J. Liu, P. Li, Y. D. Zhang, C. Guo, J. Y. Deng, J. W. Cai and B. L. Liu, “Spectroscopic Study on Binding of Folic Acid to Human Serum Albumin,” Chinese Journal of Spectroscopy Laboratory, Vol. 29, No. 7, 2009, pp. 1915-1919.

[8]   J. L. Wang, L. C. Fu, S. W. Zhou, Z. J. Chen, W. B. Lu, X. M. Ye, G. Z. Meng and S. Z. Fu, “The Interaction of Vitamin B6 with the Human Serum Albumin,” Spectroscopy and Spectral Anaiysis, Vol. 25, No. 6, 2005, pp. 912-915.

[9]   N. Zhang, C. X. Xu, Q. Wei, B. Du, R. Li, T. G. Zhang, D. Wu and Y. X. Dai, “Studies on the Interaction of DNA with Vitamin B12 Based on the Immobilization of dsDNA on Nano-Scale Hydroxyapatitle Coating,” Advanced Materials Letters, Vol. 1, No. 1, 2010, pp. 34-39.
http://dx.doi.org/10.5185/amlett.2010.3104

[10]   Z. Q. Jing, Y. H. Chi, J. Zhuang, X. Y. Bi and L. Zhou, “Mechanism Studies on the Combination Reaction between Bovine Serum Albumin and Zincon by Fluorescence Spectra,” Spectroscopy and Spectral Analysis, Vol. 27, No. 5, 2007, pp. 986-990.

[11]   J. H. Tang, S. D. Qi and X. G. Chen, “Spectroscopic Studies of the Interaction of Anticoagulant Rodennticide Diphacinone with Human Serum Albumin,” Journal of Molecular Structure, Vol. 779, No. 1-3, 2005, pp. 87-95.
http://dx.doi.org/10.1016/j.molstruc.2005.07.023

[12]   J. R. Lakowicz and G. Weber, “Quenching of Fluorescence by Oxygen, Probe for Structural Fluctuations in Macromolecules,” Biochemistry, Vol. 12, No. 21, 1973, pp. 4161-4170. http://dx.doi.org/10.1021/bi00745a020

[13]   X. Z. Feng, Z. Lin, L. J. Yang, C. Wang and C. L. Bai, “Investigation of the Interaction between Acridine Orange and Bovine Serum Albumin,” Talanta, Vol. 47, No. 5, 1998, pp. 1223-1229.
http://dx.doi.org/10.1016/S0039-9140(98)00198-2

[14]   M. Jiang, M. X. Xie, Y. Liu, X. Y. Li and X. Chen, “Spectroscopic Studies on the Interaction of Cinnamic Acid and Its Hydroxyl Derivatives with Human Serum Albumin,” Journal of Molecular Structure, Vol. 692, No. 1-3, 2004, pp. 71-80.
http://dx.doi.org/10.1016/j.molstruc.2004.01.003

[15]   T. Forster, “Modern Quantum Chemistry,” In: O. Sinaoglu, Ed., Vol. 3, Academic Press, New York, 1965.

[16]   G. Cristobal, R. Dos and D. M. Pierre, “Fluorescence Resonance Energy Transfer Spectroscopy Is a Reliable ‘Ruler’ for Measuring Structural Changes in Proteins: Dispelling the Problem of the Unknown Orientation Factor,” Journal of Structural Biology, Vol. 115, No. 2, 1995, pp. 175-185.
http://dx.doi.org/10.1006/jsbi.1995.1042

[17]   D. C. Saha, K. Ray and T. N. Misra, “Energy Transfer in Triton-X 100 Micelles: A Fluorescence Study,” Spectrochimica Acta Part A, Vol. 56, No. 4, 2000, pp. 797-801.
http://dx.doi.org/10.1016/S1386-1425(99)00169-9

 
 
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