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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.

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posted 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.

Funding

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.

History

Citation

Analytical Chemistry, 2019

Author affiliation

/Organisation/COLLEGE OF LIFE SCIENCES/School of Medicine/Department of Infection, Immunity and Inflammation

Version

  • AM (Accepted Manuscript)

Published in

Analytical Chemistry

Publisher

American Chemical Society

issn

0003-2700

Acceptance date

2019-07-01

Copyright date

2019

Publisher version

https://pubs.acs.org/doi/10.1021/acs.analchem.9b00484

Notes

The file associated with this record is under embargo until 12 months after publication, in accordance with the publisher's self-archiving policy. The full text may be available through the publisher links provided above.

Language

en

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