A general method for determining helix packing in membrane proteins in situ: Helices I and II are close to helix VII in the lactose permease of Escherichia coli

Jianhua Wu; H. Ronald Kaback

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A general method for determining helix packing in membrane proteins in situ: Helices I and II are close to helix VII in the lactose permease of Escherichia coli

机译：确定膜蛋白原位螺旋堆积的一般方法：在大肠杆菌的乳糖通透酶中，螺旋I和II与螺旋VII接近

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It was previously shown that coexpression of the lactose permease of Escherichia coli in two contiguous fragments leads to functional complementation. We demon- strate here that site-directed thiol crosslinking of coexpressed permease fragments can be used to determine helix proximity in situ without the necessity of purifying the permease. After coexpression of the six N-terminal (N_6) and six C-terminal (C_6) transmembrane helices, each with a single Cys residue, crosslinking was carried out in native membranes and as- sessed by the mobility of anti-C-terminal-reactive polypep- tides on immunoblots.

机译：先前已经证明，在两个连续片段中大肠杆菌的乳糖通透酶的共表达导致功能互补。在此我们证明，共表达通透酶片段的定点硫醇交联可用于原位确定螺旋结构，而无需纯化通透酶。共表达6个N末端（N_6）和6个C末端（C_6）跨膜螺旋后，每个螺旋都有一个Cys残基，然后在天然膜中进行交联，并根据抗C末端的迁移率进行评估。免疫印迹上的反应性多肽。

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《Proceedings of the National Academy of Sciences of the United States of America》 |1996年第25期|p.14498-14502|共5页
作者
Jianhua Wu; H. Ronald Kaback;
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收录信息美国《科学引文索引》(SCI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
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正文语种 eng
中图分类自然科学总论;
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1. Site-directed spin labeling and chemical crosslinking demonstrate that helix V is close to helices VII and VIII in the lactose permease of Escherichia coli [J] . Jianhua Wu, John Voss, Wayne L. Hubbell Proceedings of the National Academy of Sciences of the United States of America . 1996,第19期

机译：定点旋转标记和化学交联表明，螺旋V在大肠杆菌的乳糖通透酶中接近于螺旋VII和VIII。
2. Helix packing in the lactose permease of Escherichia coli determined by site-directed thiol cross-linking: helix I is close to helices V and XI. [J] . Wang Q, Kaback HR Biochemistry . 1999,第10期

机译：通过定点巯基交联确定大肠杆菌的乳糖渗透酶中的螺旋堆积：螺旋I与螺旋V和XI接近。
3. USE OF DESIGNED METAL-BINDING SITES TO STUDY HELIX PROXIMITY IN THE LACTOSE PERMEASE OF ESCHERICHIA COLI .1. PROXIMITY OF HELIX VII (ASP237 AND ASP240) WITH HELICES X (LYS319) AND XI (LYS358) [J] . He MM, Voss J, Hubbell WL, Biochemistry . 1995,第48期

机译：使用设计的金属结合位点研究大肠埃希氏菌的乳糖通透中的螺旋接近度1。 HELIX X（LYS319）和XI（LYS358）对HELIX VII（ASP237和ASP240）的邻近性
4. Effect of detergent on intramolecular helix-helix packing in a membrane-based peptide derived from CFTR [C] . Alex G.Therien, Charles M.Deber the International Peptide Symposium . 2001

机译：洗涤剂对克服膜基肽中的分子内螺旋螺旋填料的影响
5. Single deletion analysis and thiol cross -linking of a membrane transport protein: The lactose permease of Escherichia coli. [D] . Wolin, Christopher Dean. 2001

机译：膜转运蛋白的单缺失分析和巯基交联：大肠杆菌的乳糖通透酶。
6. A general method for determining helix packing in membrane proteins in situ: Helices I and II are close to helix VII in the lactose permease of Escherichia coli [O] . Jianhua Wu, H. Ronald Kaback 1996

机译：测定膜中螺旋堆积的一般方法原位蛋白质：螺旋I和II接近螺旋七在大肠杆菌中的乳糖通透性
7. Site-directed spin labeling and chemical crosslinking demonstrate that helix V is close to helices VII and VIII in the lactose permease of Escherichia coli. [O] . Wu, J, Voss, J, Hubbell, W L, 1996

机译：定点旋转标记和化学交联表明，螺旋V在大肠杆菌的乳糖通透酶中接近于螺旋VII和VIII。

A general method for determining helix packing in membrane proteins in situ: Helices I and II are close to helix VII in the lactose permease of Escherichia coli

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