一种松木蛋白酶抑制剂的纯化及结构稳定性研究[j]。集市。种子

L. A. Calderon, Humberto A. Filho, R. C. Teles, F. Medrano, C. Bloch, M. Santoro, S. Freitas
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引用次数: 7

摘要

采用一步阴离子交换色谱法从印加树粗提物中纯化了印加树胰蛋白酶抑制剂(ICTI)。集市。种子。ICTI是一个19.5 kDa的蛋白,对牛胰蛋白酶(EC 3.4.21.4) (Ki = 4.3 nM)具有显著的抑制活性。圆二色性分析显示该抑制剂为β型蛋白(β-链占40.4%;24.6%的β-turn和6.7%的α-helix)与Kunitz型抑制剂显示的性质一致。ICTI是一种在pH(1.6 ~ 10.0)范围内的热稳定蛋白,在pH 7.0时稳定性最高,Tm为70.0℃,ΔG25为48.5±0.7 kJ.mol-1。在pH值为1.6(22.5±1.2 kJ.mol-1)和pH值为10.0(31.5±1.0 kJ.mol-1)时,ΔG25的值表明在这些条件下蛋白质的结构稳定性降低。这可能是由于酸和碱侧链的pKa差异反映了折叠状态下非共价相互作用的变化。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Purification and structural stability of a trypsin inhibitor from Amazon Inga cylindrica [Vell.] Mart. seeds
Inga cylindrica Trypsin Inhibitor (ICTI) was purified as a single polypeptide chain by one step anion-exchange chromatography from a crude extract of Inga cylindrica (Vell.) Mart. seeds. ICTI is a 19.5 kDa protein presenting a remarkable inhibitory activity against bovine trypsin (EC 3.4.21.4) (Ki = 4.3 nM). Circular dichroism analysis revealed that this inhibitor is a β type protein (40.4% of β-strand; 24.6% of β-turn and 6.7% of α-helix) in accordance with properties displayed in Kunitz type inhibitors. ICTI is a thermal stable protein within a wide range of pH (1.6 to 10.0) exhibiting highest stability at pH 7.0 as indicated by Tm of 70.0 oC and ΔG25 of 48.5 ± 0.7 kJ.mol-1. The values of ΔG25 at pH 1.6 (22.5 ± 1.2 kJ.mol-1) and pH 10.0 (31.5 ± 1.0 kJ.mol-1) indicate a reduced structural stability of the protein under these conditions. This is likely to result from pKa differences of the acid and basic side chains reflecting the changes in the non-covalent interactions in the folded state.
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