生物物理学    Vol.2 No.4 (November 2014)

二硫苏糖醇抑制乳球蛋白淀粉样纤维的形成
Dithiothreitol Inhibits the Amyloid Fibril Formation of β-Lactoglobulin

作者: 马保亮 , 魏良淑 , 卢礼萍 , 吴 芳 ：南京农业大学理学院物理系，南京;

关键词: β-乳球蛋白； 二硫键； 淀粉样纤维； 透射电子显微镜； 二硫苏糖醇； β-Lactoglobulin； Disulfide Bonds； Amyloid Fibrils； Transmission Electron Microscopy； Dithiothreitol (DTT)

摘要:
淀粉样纤维与老年痴呆症、帕金森病和非神经性组织淀粉样变性病等人类疾病相关。运用ThT荧光、刚果红结合和透射电镜的方法研究了还原剂二硫苏糖醇对乳球蛋白淀粉样纤维形成的影响。实验结果表明：当乳球蛋白在中性条件和5M尿素中培养时，乳球蛋白会形成淀粉样纤维。然而当在溶液中加入二硫苏糖醇时，其淀粉样纤维的形成会受到抑制，并且这种抑制能力依赖于二硫苏糖醇的浓度。这些结果表明，二硫键在乳球蛋白淀粉样纤维形成过程中起着非常关键的作用。

Abstract: Amyloid fibrils are associated with Alzheimer’s disease, Parkinson’s disease, and non-neuropathic systemic amyloidosis. The effect of DTT on the amyloid formation of β-lactoglobulin has been ex-amined by thioflavin T binding, Congo red binding and transmission electron microscopy. The re-sults showed that the formation of the amyloid fibril took place when β-lactoglobulin was incubated in the presence of 5M urea and at neural pH. However, the formation of amyloid fibrils was inhibited when DTT was added into the solutions and the inhibitory effect is dependent on the concentrations of DTT. These results demonstrate that the disulfide bonds play important roles in the formation of amyloid fibrils.

文章引用: 马保亮 , 魏良淑 , 卢礼萍 , 吴 芳 (2014) 二硫苏糖醇抑制乳球蛋白淀粉样纤维的形成。 生物物理学， 2， 39-44. doi: 10.12677/BIPHY.2014.24005

参考文献

[1] Stefani, M. and Dobson, C.M. (2003) Protein aggregation and aggregate toxicity: New insights into protein folding, misfolding diseases and biological evolution. Journal of Molecular Medicine, 81, 678-689.

[2] Selkoe, D. (2003) Folding proteins in fatal ways. Nature, 426, 900-904.

[3] Hamada, D. and Dobson, C.M. (2002) A kinetic study of β-lactoglobulin amyloid fibril formation promoted by urea. Protein Science, 11, 2417-2426.

[4] Walsh, D., Lomakin, A., Benedek, G., Condron, M. and Teplow, D. (1997) Amyloid β-protein fibrillogenesis: Detection of a protofibrillar intermediate. Journal of Biological Chemistry, 272, 22364-22372.

[5] Dobson, C.M. (1999) Protein misfolding, evolution and disease. Trends in Biochemical Sciences, 24, 329-332.

[6] Kuwata, K., Hoshino, M., Forage, V., Era, S., Batt, C.A. and Goto, Y. (1999) Solution structure and dynamics of bovine β-lactoglobulin. Protein Science, 8, 2541-2545.

[7] Sakurai, K. and Goto, Y. (2002) Manipulating monomer-dimer equilibrium of bovine β-lactoglobulin by amino acid substitution. Journal of Biological Chemistry, 277, 25735-25740.

[8] Giurleo, J.T., He, X.L. and Talaga, D.L. (2008) β-lactoglobulin assembles into amyloid through sequential aggregated intermediates. Journal of Biological Chemistry, 381, 1332-1348.

[9] Hamada, D. and Dobson, C.M. (2002) A kinetic study of β-lactoglobulin amyloid fibril formation promoted by urea. Protein Science, 11, 2411-2426.

[10] Ma, B.L., Xie, J.B., Wei. L.S. and Li, W. (2013) Macromolecular crowding modulates the kinetics and morphology of amyloid self-assembly by β-lactoglobulin. International Journal of Biological Macromolecules, 53, 82-87.

[11] Xue, W.F., Homans, S.W. and Radford, S.E. (2008) Systematic analysis of nucleation-dependent polymerization reveals new insights into the mechanism of amyloid self-assembly. Proceedings of the National Academy of Sciences of the United States of America, 105, 8926-8931.

[12] Platt, G.W., Routledge, K.E., Homans, S.W. and Radford, S.E. (2008) Fibril growth kinetics reveal a region of β2-mi- croglobulin important for nucleation and elongation of aggregation. Journal of Molecular Biology, 378, 251-263.

[13] Ikeda, K., Okada, T., Sawada, S., Akiyoshi, K. and Matsuzaki, K. (2006) Inhibition of the formation amyloid β-protein fibrils using biocompatible nanogels as artifical chaperones. FEBS Letters, 580, 6587-6595.

[14] Krebs, M.R.H., Morozova-Roche, L.A., Daniel, K., Robinson, C.V. and Dobson, C.M. (2004) Observation of sequence specificity in the seeding of protein amyloid fibrils. Protein Science, 13, 1933-1938.

[15] Li, Y., Gong, H., Sun, Y., Yan, J., Cheng, B., Huang, K., et al. (2012) Dissecting the role of disulfide bonds on the amyloid formation of insulin. Biochemical and Biophysical Research Communications, 423, 373-378.

[16] Sarkar, N., Kumar, M. and Dubey, V.K. (2011) Effect of sodium tetrathionate on amyloid fibril: Insight into the role of disulfide bond in amyloid progression. Biochimie, 93, 962-968.

[17] Yamamoto, K., Yagi, H., Ozawa, D., Sasahara, K., Naiki, H. and Goto, Y. (2008) Thiol compounds inhibit the forma- tion of amyloid fibrils by β2-microglobulin at neutral pH. Journal of Molecular Biology, 376, 258-268.

[18] Cao, A.N., Hu, D.Y. and Lai, L.H. (2004) Formation of amyloid fibrils from fully reduced hen egg white lysozyme. Protein Science, 13, 319-324.

分享
Top