Comparison of lipid and detergent enzyme environments for identifying inhibitors of membrane-bound energy-transducing proteins

Elyse A Dunn; Gregory M Cook; Adam Heikal

doi:10.1016/j.mimet.2015.11.012

Comparison of lipid and detergent enzyme environments for identifying inhibitors of membrane-bound energy-transducing proteins

J Microbiol Methods. 2016 Jan:120:41-3. doi: 10.1016/j.mimet.2015.11.012. Epub 2015 Nov 28.

Authors

Elyse A Dunn¹, Gregory M Cook², Adam Heikal³

Affiliations

¹ Department of Microbiology and Immunology, University of Otago, Dunedin 9054, New Zealand.
² Department of Microbiology and Immunology, University of Otago, Dunedin 9054, New Zealand; Maurice Wilkins Centre for Molecular Biodiscovery, The University of Auckland, Private Bag 92019, Auckland 1042, New Zealand. Electronic address: greg.cook@otago.ac.nz.
³ Department of Microbiology and Immunology, University of Otago, Dunedin 9054, New Zealand; Maurice Wilkins Centre for Molecular Biodiscovery, The University of Auckland, Private Bag 92019, Auckland 1042, New Zealand. Electronic address: adam.heikal@otago.ac.nz.

PMID: 26593449
DOI: 10.1016/j.mimet.2015.11.012

Abstract

This study compared detergent-solubilised (soluble) and lipid-reconstituted (proteoliposome) protein to establish a high-throughput method for identifying membrane protein inhibitors. We identified inhibitors of the membrane-bound type II NADH dehydrogenase with lower lipophilicity and better potency, suggesting proteoliposome systems may be advantageous over detergent-solubilised systems for respiratory membrane proteins.

Keywords: Antibiotic; Drug discovery; High throughput screening (HTS); Proteoliposomes; Type II NADH dehydrogenase (NDH-2).

Publication types

Comparative Study
Research Support, Non-U.S. Gov't

MeSH terms

Anti-Infective Agents / pharmacology
Cell Membrane / chemistry
Cell Membrane / drug effects
Detergents / chemistry*
Detergents / pharmacology
Drug Discovery
Drug Evaluation, Preclinical
Electrophoresis, Polyacrylamide Gel
High-Throughput Screening Assays / methods
Kinetics
Lipids / chemistry*
Lipids / pharmacology
Membrane Proteins / antagonists & inhibitors*
Membrane Proteins / chemistry
Membrane Proteins / isolation & purification*
NADH Dehydrogenase / antagonists & inhibitors
NADH Dehydrogenase / chemistry
NADH Dehydrogenase / isolation & purification
Proteolipids / chemistry
Proteolipids / pharmacology

Substances

Anti-Infective Agents
Detergents
Lipids
Membrane Proteins
Proteolipids
proteoliposomes
NADH dehydrogenase II
NADH Dehydrogenase