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Cryo-EM studies of YiiP


EMSL Project ID
51299

Abstract

Cation Diffusion Facilitators are membrane transporters responsible for homeostasis of transition metals, such as Zn, Mn and Co. We are studying a bacterial homolog called YiiP, which like Znt mammalian homologs, function as Zn/H antiporters. Previous work has established the architecture of the dimeric assembly, which is distinct from other well-studied membrane transporters, three sites that bind Zn ions, and a conformational change that is consistent with the alternating access transport mechanism. Although our initial structural work relied on helical assemblies of YiiP within reconstituted lipid bilayer, we have recently produced Fab antibody fragments and generated cryo-EM structures of the complex at ~4 A resolution both in holo and apo states. We have also generated mutations that prevent Zn binding at individual sites. We now wish to generate a series of structures for these mutants to explore allosteric effects of Zn binding at each site and their roles in transport.

Project Details

Start Date
2020-03-15
End Date
2021-03-17
Status
Closed

Team

Principal Investigator

David Stokes
Institution
New York University School of Medicine

Team Members

Theo Humphreys
Institution
Oregon Health & Science University

Maria Lopez-Redondo
Institution
New York University School of Medicine

Marie Sweet
Institution
New York University School of Medicine

Related Publications

Lopez-Redondo, Maria Luisa; Koide, Akiko; Novoa-Aponte, Lorena; Fan Shujie; Beckstein, Oliver; Argüello, José M.; Koide, Shohei; Stokes, David L. (2021) Role of individual zinc binding sites in the Cation Diffusion Facilitator YiiP. 2021 Biophysical Society Meeting Abstracts. Biophysical Journal, Supplement, Abstract.