Characterization of recombinantly produced spider flagelliform silk domains
Characterization of recombinantly produced spider flagelliform silk domains
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DOI：
10.1016/j.jsb.2009.12.025
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发表时间：
2010-05-01
影响因子：
3
通讯作者：
Scheibel, Thomas
中科院分区：
文献类型：
--
作者：
Heim, Markus;Ackerschott, Christian B.;Scheibel, Thomas
蜘蛛圆网的捕获螺旋由鞭毛状的丝制成，具有高弹性和出色的韧性，非常适合捕获猎物。鞭毛状丝主要包含一种估计分子量为360 kDa的单一蛋白质（FLAG）。我们设计了模拟FLAG不同结构域的构建体（eFLAG），并重组产生它们，以分析FLAG结构域的结构-功能关系和FLAG的组装特性。虽然在溶液中小的羧基末端结构域是结构化的，但来自重复核心区域的结构域采用本质上非结构化蛋白质的典型构象。为了研究各个结构域对溶解度和组装的影响，我们测试了在已知触发丝组装的条件存在下单个结构域和结构域集合的聚集行为。重复核心结构域的长度以及羧基末端非重复结构域的存在都显示出对eFLAG聚集的影响。(C)2009 Elsevier Inc. All rights reserved.
The capture spiral of a spider's orb web is made of flagelliform silk, providing high elasticity and an outstanding toughness, perfectly suited for trapping prey. Flagelliform silk comprises mainly one single protein (FLAG) with an estimated molecular weight of 360 kDa. We engineered constructs mimicking distinct domains of FLAG (eFLAG) and produced them recombinantly to analyze the structure-function relationship of FLAG domains and assembly properties of FLAG. While in solution the small carboxy-terminal domain is structured, domains from the repetitive core region adopt a conformation typical for intrinsically unstructured proteins. To investigate the influence of the respective domains on solubility and assembly, we tested the aggregation behaviour of individual domains and domain ensembles in presence of conditions known to trigger silk assembly. Both, the length of the repetitive core domain as well as the presence of the carboxy-terminal non-repetitive domain showed impact on eFLAG aggregation. (C) 2009 Elsevier Inc. All rights reserved.