UNIT 29.9 Expression and Purification of Haemophilus influenzae Rhomboid Intramembrane Protease GlpG for Structural Studies
Published Online: 1 APR 2014
Copyright © 2013 John Wiley & Sons, Inc. All rights reserved.
Lab Protocol Title
Current Protocols in Protein Science
How to Cite
Panwar, P. and Lemieux, M. J. 2014. Expression and Purification of Haemophilus influenzae Rhomboid Intramembrane Protease GlpG for Structural Studies. Current Protocols in Protein Science. 76:29.9:29.9.1–29.9.25.
- Published Online: 1 APR 2014
Rhomboid proteases are membrane-embedded proteases that cleave peptide bonds of transmembrane proteins. They play a variety of roles in cell signaling events. The rhomboid protease GlpG from Haemophilus influenzae (hiGlpG) is a canonical form of rhomboid protease having six transmembrane segments. In this unit, detailed protocols are presented for optimization of hiGlpG expression using the araBAD promotor system in the pBAD vector. The parameters for optimization include concentration of inducing agent, induction temperature, and time. Optimization of these key factors led to the development of a protocol yielding 1.6 to 2.5 mg/liter protein purified after ion metal affinity chromatography (IMAC). Further purification can include size exclusion chromatography (SEC). Curr. Protoc. Protein Sci. 76:29.9.1-29.9.25 © 2014 by John Wiley & Sons, Inc.
- rhomboid protease;
- expression study;