Search bioRxiv⌕ Search

Biology subjects

Duerr, K. L.

Publications and source records attributed to Duerr, K. L..

2 recordsLinked to original sources

Structural Basis for Inhibition of Human Anion Transporter SLC26A6/PAT-1

PAT-1, encoded by the gene SLC26A6, is an exchanger of small organic and inorganic anions including oxalate and chloride in the kidney and the intestines. PAT-1 dysfunction is implicated in nephrolithiasis and hyperoxaluria, and its modulation is a potential therapeutic strategy to treat cystic fibrosis. PAT-1 has two small molecule inhibitors: DIDS, a non-specific anion exchanger inhibitor; and PAT1inh-B01 which is highly specific to PAT-1. In this study, we report high-resolution cryo-electron microscopy structures of the human PAT-1 homodimer in complex with these inhibitors to reveal their different inhibition mechanisms. These structures also uncover the complex interactions between the transmembrane, STAS and N-terminal domains, with implications on its ion exchange function. Hence, this study provides mechanistic insight into PAT-1 molecular function and inhibition, and contributes to our understanding of SLC26 transport mechanisms.

biochemistry↗

Di-Gluebodies as covalently-rigidified, modular protein assemblies enable simultaneous determination of high-resolution, low-size, cryo-EM structures

Cryo-EM has become a routine structural biology method, yet elucidation of small proteins (<100 kDa) and increasing throughput remain challenging. Here, we describe covalently-dimerized engineered nanobodies, Di-Gluebodies, as novel and modular imaging scaffolds to address the challenges. They can simultaneously display two copies of the same protein or two distinct proteins, providing sufficient rigidity required by cryo-EM. We validate this method with multiple soluble and membrane targets, demonstrating that this provides a flexible and scalable platform for expanded protein structure determination.

synthetic biology↗