The high-throughput production of membrane proteins

DOI: 10.1042/ETLS20210196

Authors: James Birch (Diamond Light Source; Research Complex at Harwell (RCaH)) , Andrew Quigley (Diamond Light Source; Research Complex at Harwell (RCaH))
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Emerging Topics In Life Sciences , VOL 6

State: Published (Approved)
Published: October 2021

Abstract: Membrane proteins, found at the junctions between the outside world and the inner workings of the cell, play important roles in human disease and are used as biosensors. More than half of all therapeutics directly affect membrane protein function while nanopores enable DNA sequencing. The structural and functional characterisation of membrane proteins is therefore crucial. However, low levels of naturally abundant protein and the hydrophobic nature of membrane proteins makes production difficult. To maximise success, high-throughput strategies were developed that rely upon simple screens to identify successful constructs and rapidly exclude those unlikely to work. Parameters that affect production such as expression host, membrane protein origin, expression vector, fusion-tags, encapsulation reagent and solvent composition are screened in parallel. In this way, constructs with divergent requirements can be produced for a variety of structural applications. As structural techniques advance, sample requirements will change. Single-particle cryo-electron microscopy requires less protein than crystallography and as cryo-electron tomography and time-resolved serial crystallography are developed new sample production requirements will evolve. Here we discuss different methods used for the high-throughput production of membrane proteins for structural biology.

Journal Keywords: high-throughput screening; protein expression; protein purification; structural biology; transmembrane proteins

Subject Areas: Biology and Bio-materials

Diamond Offline Facilities: Membrane Protein Laboratory (MPL)
Technical Areas:

Added On: 12/10/2021 09:40

Documents:
etls-2021-0196c.pdf

Discipline Tags:

Structural biology Life Sciences & Biotech

Technical Tags: