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首页> 外文期刊>Journal of Molecular Biology >A compact monomeric intermediate identified by NMR in the denaturation of dimeric triose phosphate isomerase.
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A compact monomeric intermediate identified by NMR in the denaturation of dimeric triose phosphate isomerase.

机译：在二聚磷酸三糖磷酸异构酶变性中通过NMR鉴定的致密单体中间体。

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摘要

The denaturation of triose phosphate isomerase (TIM) from Saccharomyces cerevisiae by guanidine hydrochlorids at pH 7.2 has been monitored by NMR spectroscopy in conjunction with optical spectroscopy. In the absence of denaturant, the hydrodynamic radius of 29.6(+/-0.25) A and the substantial chemical shift dispersion evident in the NMR spectrum are consistent with the highly structured dimeric native state of the protein. On the addition of 2. 2 M guanidine hydrochloride the effective hydrodynamic radius increases to 51.4(+/-0.43) A, consistent with that anticipated for the polypeptide chain in a highly unstructured random coil state. In 1.1 M guanidine hydrochloride, however, the effective hydrodynamic radius is 24.0(+/-0.25) A, a value substantially decreased relative to that of the native dimeric state but very close to that anticipated for a monomeric species with native-like compaction (23. 5 A). The lack of chemical shift dispersion indicates, however, that few tertiary interactions persist within this species. Far UV CD and intrinsic fluorescence measurements show that this compact intermediate retains significant secondary structure and that on average the fluorophores are partially excluded from solvent. Such a species could be important in the formation of dimeric TIM from its unfolded state. Copyright 2000 Academic Press.
机译：已经通过NMR光谱法和光学光谱法监测了在pH 7.2下盐酸胍对啤酒酵母中的磷酸三糖异构酶（TIM）的变性。在没有变性剂的情况下，NMR谱图中明显的29.6（+/- 0.25）A的流体力学半径和明显的化学位移分散与蛋白质的高度结构化的二聚体天然状态相符。加入2. 2 M盐酸胍后，有效流体力学半径增加至51.4（+/- 0.43）A，与高度无规无规卷曲状态下多肽链的预期流体动力学半径一致。然而，在1.1 M盐酸胍中，有效流体力学半径为24.0（+/- 0.25）A，相对于天然二聚体状态而言，该值显着降低，但与具有天然样致密性的单体物种的预期值非常接近（ 23. 5 A）。然而，缺乏化学位移分散性表明该物种内几乎没有三级相互作用。远紫外CD和固有荧光测量表明，这种紧凑的中间体保留了重要的二级结构，并且平均而言，荧光团被部分排除在溶剂之外。这样的物种对于从其展开状态形成二聚体TIM可能是重要的。版权所有2000学术出版社。

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