Mechanical unfoldons as building blocks of maltose-binding protein

被引:61
作者
Bertz, Morten [1 ]
Rief, Matthias [1 ,2 ]
机构
[1] Tech Univ Munich, Phys Dept E22, D-85748 Garching, Germany
[2] Munich Ctr Integrated Protein Sci CiPSM, Munich, Germany
关键词
single-molecule force spectroscopy; protein folding; energy landscape; unfolding intermediate; atomic force microscopy;
D O I
10.1016/j.jmb.2008.02.025
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
Identifying independently folding cores or substructures is important for understanding and assaying the structure, function and assembly of large proteins. Here, we suggest mechanical stability as a criterion to identify building blocks of the 366 amino acid maltose-binding protein (MBP). We find that MBP, when pulled at its termini, unfolds via three (meta-) stable unfolding intermediates. Consequently, the MBP structure consists of four structural blocks (unfoldons) that detach sequentially from the folded structure upon force application. We used cysteine cross-link mutations to characterize the four unfoldons structurally. We showed that many MBP constructs composed of those building blocks indeed form stably folded structures in solution. Mechanical unfoldons may provide a new tool for a systematic search for stable substructures of large proteins. (c) 2008 Elsevier Ltd. All rights reserved.
引用
收藏
页码:447 / 458
页数:12
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