Conformational properties of the aggregation precursor state of HypF-N

被引:42
作者
Campioni, Silvia [1 ]
Mossuto, Maria F. [2 ]
Torrassa, Silvia [3 ]
Calloni, Giulia [1 ]
de Laureto, Patrizia Polverino [2 ]
Relini, Annalisa [3 ]
Fontana, Angelo [2 ]
Chiti, Fabrizio [1 ]
机构
[1] Univ Florence, Dipartimento Sci Biochim, I-50134 Florence, Italy
[2] Univ Padua, CRIBI Biotechnol Ctr, I-35121 Padua, Italy
[3] Univ Genoa, Dipartimento Fis, I-16146 Genoa, Italy
关键词
acid-denatured state; pre-molten globule; amyloid fibrils; amyloidogenic state; protein aggregation;
D O I
10.1016/j.jmb.2008.04.002
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The conversion of specific proteins or protein fragments into insoluble, ordered fibrillar aggregates is a fundamental process in protein chemistry, biology, medicine and biotechnology. As this structural conversion seems to be a property shared by many proteins, understanding the mechanism of this process will be of extreme importance. Here we present a structural characterisation of a conformational state populated at low pH by the N-terminal domain of Escherichia coli HypE Combining different biophysical and biochemical techniques, including near- and far-UV circular dichroism, intrinsic and 8-anilinonaphthalene-1-sulfonate-derived fluorescence, dynamic light scattering and limited proteolysis, we will show that this state is largely unfolded but contains significant secondary structure and hydrophobic clusters. It also appears to be more compact than a random coil-like state but less organised than a molten globule state. Increase of the total ionic strength of the solution induces aggregation of such a pre-molten globule state into amyloid-like protofibrils, as revealed by thioflavin T fluorescence and atomic force microscopy. These results show that a premolten globule state can be, among other possible conformational states, one of the precursor states of amyloid formation. In addition, the possibility of triggering aggregation by modulating the ionic strength of the solution provides one a unique opportunity to study both the initial precursor state and the aggregation process. (c) 2008 Elsevier Ltd. All rights reserved.
引用
收藏
页码:554 / 567
页数:14
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