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Phospho-regulation, nucleotide binding and ion access control in potassium-chloride cotransporters

EMBO J. 2021-05; 
Gamma Chi, Rebecca Ebenhoch, Henry Man, Haiping Tang, Laurence E Tremblay, Gabriella Reggiano, Xingyu Qiu, Tina Bohstedt, Idlir Liko, Fernando G Almeida, Alexandre P Garneau, Dong Wang, Gavin McKinley, Christophe P Moreau, Kiran D Bountra, Patrizia Abrusci, Shubhashish M M Mukhopadhyay, Alejandra Fernandez-Cid, Samira Slimani, Julie L Lavoie, Nicola A Burgess-Brown, Ben Tehan, Frank DiMaio, Ali Jazayeri, Paul Isenring, Carol V Robinson, Katharina L Dürr
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GenCrispr Cas9 Genome Editing Full‐length NKCC1, phospho‐mimetic (S45D, T940D, T997D) and phospho‐knockout (S45A, T940A, T997A) constructs of KCC3b were synthesized (GenScript, Twist Bioscience)? Get A Quote

摘要

Potassium-coupled chloride transporters (KCCs) play crucial roles in regulating cell volume and intracellular chloride concentration. They are characteristically inhibited under isotonic conditions via phospho-regulatory sites located within the cytoplasmic termini. Decreased inhibitory phosphorylation in response to hypotonic cell swelling stimulates transport activity, and dysfunction of this regulatory process has been associated with various human diseases. Here, we present cryo-EM structures of human KCC3b and KCC1, revealing structural determinants for phospho-regulation in both N- and C-termini. We show that phospho-mimetic KCC3b is arrested in an inward-facing state in which intracellular ion access is ... More

關鍵詞

HDX-MS, nucleotide binding, phospho-regulation, potassium-chloride co-transport, solute carrier
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