High-Throughput Mass Spectrometry Applied to Structural Genomics

Chromatography

Rod Chalk¹  Georgina Berridge¹  Leela Shrestha²  Claire Strain-Damerell²  Pravin Mahajan²  Wyatt W. Yue²  Opher Gileadi²  Nicola A. Burgess-Brown¹ 

[1] id="af1-chromatography-01-00159">Structural Genomics Consortium, University of Oxford, Old Road Campus Research Building, Roosevelt Drive, Oxford OX3 7DQ,

关键词: high-throughput;    mass spectrometry;    protein expression;    structural genomics;    integral membrane protein (IMP);    post-translational modification (PTM);    intact mass;   

DOI  :  10.3390/chromatography1040159

来源: mdpi

【 摘 要 】

Mass spectrometry (MS) remains under-utilized for the analysis of expressed proteins because it is inaccessible to the non-specialist, and sample-turnaround from service labs is slow. Here, we describe 3.5 min Liquid-Chromatography (LC)-MS and 16 min LC-MSMS methods which are tailored to validation and characterization of recombinant proteins in a high throughput structural biology pipeline. We illustrate the type and scope of MS data typically obtained from a 96-well expression and purification test for both soluble and integral membrane proteins (IMPs), and describe their utility in the selection of constructs for scale-up structural work, leading to cost and efficiency savings. We propose that value of MS data lies in how quickly it becomes available and that this can fundamentally change the way in which it is used.

【 授权许可】

CC BY   
© 2014 by the authors; licensee MDPI, Basel, Switzerland.