6ADA

Crystal structure of the E148D mutant CLC-ec1 in 200mM bromide

6ADA の概要

• エントリーDOI: 10.2210/pdb6ada/pdb
• 分子名称: H(+)/Cl(-) exchange transporter ClcA, antibody Fab fragment, heavy chain, antibody Fab fragment, light chain, ... (4 entities in total)
• 機能のキーワード: clc cl-/h+ antiporter, intermediate structure, external glutamate, transport protein
• 由来する生物種: Escherichia coli (strain K12); 詳細
• タンパク質・核酸の鎖数: 6
• 化学式量合計: 194895.31

構造登録者

Lim, H.-H.,Park, K. (登録日: 2018-07-31, 公開日: 2019-08-28, 最終更新日: 2024-10-16)

主引用文献

Park, K.,Lee, B.C.,Lim, H.H.
Mutation of external glutamate residue reveals a new intermediate transport state and anion binding site in a CLC Cl-/H+antiporter.
Proc.Natl.Acad.Sci.USA, 116:17345-17354, 2019

PubMed Abstract: The CLC family of proteins are involved in a variety of physiological processes to control cellular chloride concentration. Two distinct classes of CLC proteins, Cl channels and Cl/H antiporters, have been functionally and structurally investigated over the last several decades. Previous studies have suggested that the conformational heterogeneity of the critical glutamate residue, Glu, could explain the transport cycle of CLC-type Cl/H antiporters. However, the presence of multiple conformations (, , and ) of the Glu has been suggested from combined structural snapshots of 2 different CLC antiporters: CLC-ec1 from and cmCLC from a thermophilic red alga, Thus, we aimed to investigate further the heterogeneity of Glu-conformations in CLC-ec1, the most deeply studied CLC antiporter, at both functional and structural levels. Here, we show that the crystal structures of the Glu mutant E148D and wild-type CLC-ec1 with varying anion concentrations suggest a structural intermediate, the "" conformation. We also found that an extra anion can be located above the external Cl-binding site in the E148D mutant when the anion concentration is high. Moreover, we observed that a carboxylate in solution can occupy either the external or central Cl-binding site in the ungated E148A mutant using an anomalously detectable short carboxylic acid, bromoacetate. These results lend credibility to the idea that the Glu can take at least 3 distinct conformational states during the transport cycle of a single CLC antiporter.

PubMed: 31409705
DOI: 10.1073/pnas.1901822116

実験手法

X-RAY DIFFRACTION (3.153 Å)