Membrane Proteins

A Typical Membrane Protein Screening and Production Cascade

Working with membrane proteins has a high degree of difficulty, and they are one of the most challenging types of target that we work with. There are many steps from the expression through the purification that can require extra problem solving or optimisation. We take an approach that will allow the collection of as much feedback on how to optimise the process as early as possible and create opportunities to communicate with you throughout the process to discuss data and explain our recommendations for the next steps. We aim to drive clear, data driven decision points between stages to help manage costs.

Membrane Proteins Process Overview

Construct Design
Construct Design

Construct Design

In collaboration with you, and based on literature review, modelling, in-house software and any prior in-house experience, we will design a set of systematic constructs that best suit your needs. Typical variations include exploration of purification tags, stabilising fusions and stabilising point mutations.

Protein Expression
Protein Expression

Expression and Solubilisation Screening

In initial testing, small-scale expression studies are typically carried out to screen expression host, protein construct and solubilisation detergent with the aim to define the optimal combination. Typically, we use FSEC and Western blot analysis to help drive our selection criteria.

Protein Purification
Protein Purification

Feasibility Purifications

A selection of key candidates from previous stage are typically scaled-up to 1 L expression cultures, and feasibility purifications are carried out. With these purifications, we aim to confirm yields following purification, quality and perform optimisations of the process. A outcome is typically to define the final presentation of the sample (e.g. detergent, nanodisc, liposome).

Protein Characterisation
Protein Characterisation

Scale Up

Once conditions have been optimised, we will scale-up the expression and purification to produce your membrane protein in sufficient quantity and with excellent stability and quality characteristics for your end use.

Cryo-EM Full Data Collection

Validation of Protein Function and Quality

If required, additional quality control and ligand binding experiments can be carried out to assess the protein we have produced for you. We can use biophysical assays such as FIDA, SEC-MALS and thermostability shift stability assays (TSA) to perform these experiments.

Hear from one of our experts

Mark Elvin: Head of Cell Science

“Our specialist team of membrane protein experts have extensive experience tackling some of the most challenging targets. Production of high-quality membrane protein reagents allow us to drive transformative insights and solutions in drug discovery and biotechnology for our clients.”

Steve Harborne: Head of Membrane Protein Science

We have successfully produced membrane proteins from the following classes

GPCR’s

Membrane Proteins - GPCRs

Solute Transporters

Membrane Proteins -

ION Channels

Membrane Proteins - ION Channels

GPCR:G-Protein Complexes

Membrane Proteins - GPCR:G-Porotein Complexes

ABC Transporters

Membrane Proteins - ABC Transporters

Receptors

Membrane Proteins - Receptors

Purified Membrane Proteins Can Be Presented in Different Formats

Detergent

Detergent

SMALP

SMALP

Nanodisc

Nanodisc

Liposome

Liposome

Why Choose Peak Proteins for your Membrane Protein Project

A dedicated team with expertise and a strong track record in membrane protein production

Experience with membrane protein crystallization in both vapour diffusion and lipidic cubic phase experiments

Experience with preparing membrane proteins for cryo-EM

Equipment and Software Specific for membrane proteins.

  • IMPROvER software for membrane protein stability.
  • Ultracentrifuges (Beckman Optima benchtop and floor standing)
  • LCP capable crystallisation mosquito

Membrane Proteins Resources

TitleContentCategoriesResources Typeshf:categorieshf:tax:resource_type
Determining the Cryo-EM Structure of the 5-HT2A receptor with Serotonin BoundCase StudyMOREcase-study
Purifying 5-HT2AR GPCR for cryo-EM Structural Studies, , BlogMOREmembrane-proteins blog peak-favouritesblog
Using the BacMam System in HEK293-6E cells to Generate High Level Expression of a Membrane ProteinCase StudyMOREcase-study
Introduction to Detergents for Membrane Protein Solubilisation, , BlogMOREmembrane-proteins blog peak-favouritesblog
Applications of FIDA in Protein Science, , PostersMOREfida membrane-proteins protein-purificationposters
Using the Power of Cryo-EM for Structure Determination at Peak Proteins, , , BlogMOREcryo-em membrane-proteins blog protein-purificationblog
Rapid Assessment of Membrane Protein Quality by Fluorescent Size Exclusion Chromatography, , , MOREpeak-protein-news membrane-proteins peak-favourites protein-purification
Protein Solubility Predictor ToolThe protein-sol software will take a single amino acid sequence and return the result of a set of solubility prediction …, LinksMOREmembrane-proteins protein-purificationlinks
IMPROvER ToolIntegral Membrane PROtein stability sElectoR. Software that uses three different approaches to identify and rank potentially stabilising mutations in α-helical membrane proteins., LinksMOREmembrane-proteins protein-purificationlinks