The Bacteroides dual-pumping membrane-integral pyrophosphatase: a novel drug target

Información del proyecto

BactoDrug

Identificador del acuerdo de subvención: 659677

DOI

10.3030/659677

Proyecto finalizado el 30 Abril 2017

Fecha de la firma de la CE 23 Marzo 2015

Fecha de inicio 1 Junio 2015

Fecha de finalización 31 Mayo 2017

Financiado con arreglo a

• EXCELLENT SCIENCE - Marie Skłodowska-Curie Actions

Coste total

€ 183 454,80

Aportación de la UE

€ 183 454,80

Coordinado por

UNIVERSITY OF LEEDS
United Kingdom

Objetivo

Membrane-integral pyrophosphatases (M-PPases) couple cleavage of pyrophosphatase to pumping of ions across a membrane to generate membrane potential and play an important role in resistance to stressors. The solved structures of an H+-pumping M-PPase from Vigna radiata and an Na+-pumping M-PPase from Thermotoga maritima show M-PPases form a channel through the membrane, and this channel is plugged by an ion gate formed by three charged residues. Despite these structures, there are still many outstanding questions regarding M-PPases, especially in relation to H+ and Na+ dual-pumping M-PPases.

Bacteroides species are a major cause of anaerobic infections, and though they are part of a healthy human gut flora, when these bacteria escape the gut, they can cause bacteremia and abscess formation. Bacteroides species are associated with high antibiotic resistance rates and have a 19% or greater mortality rate. However, they do possess a possible drug target: an H+/Na+-pumping M-PPase.

A major goal of this project is to solve the structure of the Bacteroides vulgatus H+/Na+-pumping M-PPase to guide mutational studies to determine how M-PPases select for ions and to explore how the ion gate is opened and closed during ion pumping. Since the ion gate is closed in all M-PPase structures to date, I will also use single molecule fluorescence resonance energy transfer and total internal reflection fluorescence microscopy to determine the kinetics and conformational changes during ion gate movement. Finally, I will use molecular mechanics modeling to simulate ion gate function and design small-molecule drug candidates. Molecules that trap the ion gate in the open conformation will convert M-PPase into a pore in the membrane of Bacteroides species, leading to collapse of the membrane potential.

This project will further my career goal of pursuing research in bacterial pathogenesis from various perspectives, utilizing X-ray crystallography and single molecule technologies.

Ámbito científico (EuroSciVoc)

• ciencias naturales › ciencias biológicas › microbiología › bacteriología
• ciencias naturales › ciencias de la tierra y ciencias ambientales conexas › geología › mineralogía › cristalografía
• ciencias naturales › ciencias físicas › óptica › microscopía
• ciencias naturales › ciencias biológicas › bioquímica › biomoléculas › lípidos
• ciencias naturales › ciencias biológicas › bioquímica › biomoléculas › proteínas › enzima

Palabras clave

• integral membrane protein
• X-ray crystallography
• membrane-integral pyrophosphatase

Programa(s)

• H2020-EU.1.3. - EXCELLENT SCIENCE - Marie Skłodowska-Curie Actions PROGRAMA PRINCIPAL
  
• H2020-EU.1.3.2. - Nurturing excellence by means of cross-border and cross-sector mobility
  

Tema(s)

• MSCA-IF-2014-EF - Marie Skłodowska-Curie Individual Fellowships (IF-EF)
  

Régimen de financiación

MSCA-IF - Marie Skłodowska-Curie Individual Fellowships (IF)

Convocatoria de propuestas

H2020-MSCA-IF-2014

Coordinador

UNIVERSITY OF LEEDS

• Aportación neta de la UEn: € 183 454,80
• Dirección: WOODHOUSE LANE ; LS2 9JT Leeds ; Reino Unido
• Región: Yorkshire and the Humber › West Yorkshire › Leeds
• Tipo de actividad: Higher or Secondary Education Establishments
• Coste total: € 183 454,80