Loraine Alhumaidan 2019 Analytical Chemistry.pdf (808.2 kB)
Efficient Protein Digestion at Elevated Temperature in the Presence of SDS and Calcium Ions for Membrane Proteomics.
journal contributionposted on 2019-07-08, 15:30 authored by J Loraine, O Alhumaidan, A Bottrill, S Mistry, P Andrew, G Mukamolova, O Turapov
Growing significance of membrane proteins inspires continuous development and improvement of methods for robust membrane proteomics. Here, we developed a very simple and efficient method for membrane protein digestion using an ionic detergent SDS at high temperature, conditions where trypsin is normally inactivated. Our results suggest that trypsin can be stabilized by a combination of calcium ions and sodium chloride which enables protein digestion at elevated temperature in the presence of SDS. Application of this method resulted in higher efficiency of digestion and improved identification of membrane proteins, offering novel opportunities for advancement in membrane proteomics.
The project was supported by the UK Biotechnology and Biological Sciences Research Council grants BB/H008586/1 and BB/K000330/1 (to G.V.M.), MRC-DTG (to J.L.). Ministry of Education in Saudi Arabia. Riyadh, Saudi Arabia.
CitationAnalytical Chemistry, 2019
Author affiliation/Organisation/COLLEGE OF LIFE SCIENCES/School of Medicine/Department of Infection, Immunity and Inflammation
- AM (Accepted Manuscript)