Decoding the folding of Burkholderia glumae lipase: folding intermediates en route to kinetic stability.
Decoding the folding of Burkholderia glumae lipase: folding intermediates en route to kinetic stability.
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解码Burkholderia glumae脂肪酶的折叠：在动力学稳定性的途中折叠中间体。
DOI：
10.1371/journal.pone.0036999
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Van Gelder P
中科院分区：
文献类型：
--
作者：
Pauwels K;Sanchez del Pino MM;Feller G;Van Gelder P
由Burkholderia glumae产生的脂肪酶自发折叠成非活性的近天然状态，并且需要周质伴侣来达到其最终的活性和分泌能力折叠。B。颖壳脂肪酶特异性折叠酶（Lif）属于空间伴侣蛋白家族，其中α-裂解蛋白酶和枯草杆菌蛋白酶的前肽是最著名的代表。空间伴侣在赋予蛋白质动力学稳定性方面起关键作用。然而，到目前为止，没有确凿的实验证据表明，生命依赖性脂肪酶是动力学捕获的酶。通过结合热变性研究与蛋白水解抗性实验和不同的折叠中间体的描述，我们表明，天然脂肪酶具有动力学稳定的构象。我们表明，一个新发现的熔融球状构象具有明显不同于那些近天然的中间态的独特属性。的折叠指纹的生命依赖性脂肪酶的蛋白酶前结构域系统的上下文中，比较揭示了明确的差异，使脂肪酶，Lif系统的独特。有限的蛋白水解揭示了近天然中间体和天然构象之间的结构差异，并为阐明动力学屏障的性质奠定了基础。
The lipase produced by Burkholderia glumae folds spontaneously into an inactive near-native state and requires a periplasmic chaperone to reach its final active and secretion-competent fold. The B. glumae lipase-specific foldase (Lif) is classified as a member of the steric-chaperone family of which the propeptides of α-lytic protease and subtilisin are the best known representatives. Steric chaperones play a key role in conferring kinetic stability to proteins. However, until present there was no solid experimental evidence that Lif-dependent lipases are kinetically trapped enzymes. By combining thermal denaturation studies with proteolytic resistance experiments and the description of distinct folding intermediates, we demonstrate that the native lipase has a kinetically stable conformation. We show that a newly discovered molten globule-like conformation has distinct properties that clearly differ from those of the near-native intermediate state. The folding fingerprint of Lif-dependent lipases is put in the context of the protease-prodomain system and the comparison reveals clear differences that render the lipase-Lif systems unique. Limited proteolysis unveils structural differences between the near-native intermediate and the native conformation and sets the stage to shed light onto the nature of the kinetic barrier.
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DOI：
10.1107/s1744309105023055
发表时间：
2005-08-01
影响因子：
0.9
作者：
Pauwels, K;Loris, R;Van Gelder, P
通讯作者：
Van Gelder, P
影响因子：
2.9
作者：
LEPOCK, JR;RITCHIE, KP;KRUUV, J
通讯作者：
KRUUV, J
DOI：
10.1093/protein/11.5.349
发表时间：
1998-05-01
期刊：
PROTEIN ENGINEERING
影响因子：
--
作者：
Hubbard, SJ;Beynon, RJ;Thornton, JM
通讯作者：
Thornton, JM
影响因子：
5.6
作者：
Rodriguez-Larrea, David;Minning, Stefan;Sanchez-Ruiz, Jose M.
通讯作者：
Sanchez-Ruiz, Jose M.
影响因子：
4.7
作者：
Drepper, Thomas;Eggert, Thorsten;Jaeger, Karl-Erich
通讯作者：
Jaeger, Karl-Erich