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首页> 外文期刊>Journal of Molecular Biology >Helix packing in the lactose permease of Escherichia coli: Localization of helix VI
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Helix packing in the lactose permease of Escherichia coli: Localization of helix VI

机译：大肠杆菌乳糖渗透酶中的螺旋填料：螺旋VI的定位

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摘要

Plasmids encoding "split" lactose permease constructs with discontinuities in either the periplasmic loop between helices V and VI (N-5/C-7) or between helices VI and VII (N-6/C-6) were used to localize helix VI within the tertiary structure by site-directed thiol cross-linking. A total of 57 double-Cys pairs, with one Cys residue in helix VI and another in helix V or VIII, were studied with homobifunctional cross-linking agents. Significant cross-linking is observed between the periplasmic ends of helices V (position 158 or 161) and VI (position 170) with rigid 6 or 10 Angstrom reagents. Furthermore, the Cys residue at position 170 (helix VI) also cross-links to a Cys residue at either position 264 or 265 (helix VIII) with a 21 Angstrom cross-linking agent. The data indicate that helices V, VI and VIII are in close proximity at the periplasmic face of the membrane, with helix VI significantly closer to helix V. In addition, beta ,D-galactopyranosyl 1-thio-beta ,D-galactopyranoside induces a significant increase in cross-linking efficiency between helices VI and VIII and between helices V and VIII, with no significant change in cross-linking between helices V and VI.
机译：编码“分裂”乳糖通透酶构建体的质粒在螺旋V和VI之间（N-5 / C-7）或螺旋VI和VII之间（N-6 / C-6）的周质环中具有不连续性，用于定位螺旋VI在三级结构中通过定点硫醇交联。用同双功能交联剂研究了总共57个双Cys对，在螺旋VI中具有一个Cys残基，在螺旋V或VIII中具有另一个。在使用刚性6或10埃试剂的螺旋V（位置158或161）和VI（位置170）的周质末端之间观察到明显的交联。此外，在第170位（螺旋VI）处的Cys残基也用21埃交联剂与在第264或265位（螺旋VIII）处的Cys残基交联。数据表明，螺旋V，VI和VIII在膜的周质面非常接近，螺旋VI显着靠近螺旋V。此外，β，D-吡喃半乳糖基1-硫代-β，D-吡喃半乳糖苷可诱导螺旋VI和VIII之间以及螺旋V和VIII之间的交联效率显着提高，而螺旋V和VI之间的交联没有显着变化。

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