Extracellular loop 4 of the proline transporter PutP controls the periplasmic entrance to ligand binding sites.

Extracellular loop 4 of the proline transporter PutP controls the periplasmic entrance to ligand binding sites.
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脯氨酸转运蛋白 PutP 的细胞外环 4 控制配体结合位点的周质入口

DOI:
10.1016/j.str.2014.03.011
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发表时间:
2014
期刊:
影响因子:
5.7
通讯作者:
Steinhoff
Steinhoff
中科院分区:
生物学2区
文献类型:
--
作者:
Dunkel;Hilger;Lipiszko;Polyhach;Jeschke;Bracher;Steinhoff

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Na+/脯氨酸同向转运体(PutP)与其他几种Na+偶联同向转运体一样,属于所谓的LeuT折叠结构家族,其特征在于由细胞内外环连接的10个核心跨膜结构域(cTM)。这些循环的作用已经在二级主动运输过程的交替访问模型中的门控功能的上下文中进行了讨论。在这里,我们报告了PutP细胞外环4(eL 4)的完整自旋标记位点扫描,揭示了两个α螺旋片段eL 4a和eL 4 b的存在。在与转运蛋白的功能动力学直接相关的eL 4残基中,eL 4 b中的Phe 314通过与cTM 1的疏水接触(靠近配体结合位点)锚定环。我们提出,配体诱导的构象变化在结合位点通过锚定残基传递到eL 4,并通过eL 4进一步传递到相邻的cTM,导致细胞外闸门的关闭。
The Na+/proline symporter (PutP), like several other Na+-coupled symporters, belongs to the so-called LeuT-fold structural family, which features ten core transmembrane domains (cTMs) connected by extra- and intracellular loops. The role of these loops has been discussed in context with the gating function in the alternating access model of secondary active transport processes. Here we report the complete spin-labeling site scan of extracellular loop 4 (eL4) in PutP that reveals the presence of two α-helical segments, eL4a and eL4b. Among the eL4 residues that are directly implicated in the functional dynamics of the transporter, Phe314 in eL4b anchors the loop by means of hydrophobic contacts to cTM1 close to the ligand binding sites. We propose that ligand-induced conformational changes at the binding sites are transmitted via the anchoring residue to eL4 and through eL4 further to adjacent cTMs, leading to closure of the extracellular gate.
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