Detergent Screen for solubilized membrane proteins – Case study on the SLAC-protein HiTehA from Haemophilus influenzae

November 9, 2018

The biophysical characterization of integral membrane protein stability is often challenging due to several factors: First, the expression and purification of membrane proteins is often impeded by low expression levels and protein stability. As a result, yields are usually low and do not allow for a thorough analysis or a screening approach to determine optimal conditions. Second, the use of detergents – which are necessary to solubilize membrane proteins – often introduces artifacts and other secondary effects, and most importantly precludes the use of reporter dyes to monitor protein unfolding. Label free methods – such as DSC or CD spectroscopy – on the other hand require large quantities of proteins, and are limited in throughput. 

Here we use the 10 transmembrane-helix protein HiTehA, a protein of the slow anion channel family, to present label-free, native DSF as the method of choice to perform rapid and precise detergent screening projects for a solubilized membrane protein.

Previous Article
High-throughput stability screening for detergent-solubilized membrane proteins
High-throughput stability screening for detergent-solubilized membrane proteins

No More Flipbooks

Working with tricky membrane proteins?

NanoTemper tools can help

Find out how