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首页> 外文期刊>International Journal of Biological Macromolecules: Structure, Function and Interactions >Determination of structural and thermodynamic parameters of bovine alpha-trypsin isoform in aqueous-organic media
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Determination of structural and thermodynamic parameters of bovine alpha-trypsin isoform in aqueous-organic media

机译:牛α-胰蛋白酶同种型在水性有机培养基中的结构和热力学参数的测定

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摘要

The alpha-trypsin isoform is a globular protein that belongs to serine-protease family and has a polypeptide chain of 223 amino acid residues, six disulfide bridges and two domains with similar structures. The effects of aqueous-organic solvent (ethanol) in different concentration on the alpha-trypsin structure have been investigated by spectroscopic techniques and thermodynamic data analysis. The results from spectroscopic measurements, including far-UV Circular Dichroism, UV-vis absorption spectroscopy, intrinsic tryptophan fluorescence and dynamic light scattering (DLS) suggest the formation of partially folded states, instead of aggregate states, at high ethanol concentration (>60% v/v ethanol), with little loss of secondary structure, but with significant tertiary structure changes. The thermodynamic data (T-m and Delta H) suggest a loosening of intramolecular weak interactions, which reflects in a flexibility increase such that the catalytic capacity can be increased or decreased according to the ethanol concentration into the system. Overall results we suggest that in range of 0-60% v/v ethanol/buffer, alpha-trypsin undergoes reversible multimerization phenomena with catalytic activity. However from 60% v/v ethanol/buffer, population of folded partially states with less catalytic activity are predominant. (C) 2017 Elsevier B.V. All rights reserved.
机译:α-胰蛋白酶同种型是属于丝氨酸蛋白酶家族的球状蛋白质,并且具有223个氨基酸残基,六个二硫键和具有相似结构的两个结构域的多肽链。通过光谱技术和热力学数据分析研究了不同浓度对α-胰蛋白酶结构的不同浓度的效果。光谱测量结果,包括远UV圆形二色性,UV-Vis吸收光谱,内在色氨酸荧光和动态光散射(DLS)表明,在高乙醇浓度下形成部分折叠状态,而不是聚集状态(> 60%) v / v乙醇),缺少二级结构,但具有大量的三级结构变化。热力学数据(T-M和DELTA H)表明对分子内弱相互作用的松动,其反映了柔韧性增加,使得催化力可以根据乙醇浓度增加或减少到系统中。总体结果表明,范围为0-60%v / v乙醇/缓冲液,α-胰蛋白酶经历可逆的多聚化现象,具有催化活性。然而,从60%v / v乙醇/缓冲液中,折叠部分状态具有较少的催化活性的群体是主要的。 (c)2017年Elsevier B.V.保留所有权利。

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