UNIT 29.8 Membrane Transport Piece by Piece: Production of Transmembrane Peptides for Structural and Functional Studies
Published Online: 3 FEB 2014
Copyright © 2013 John Wiley & Sons, Inc. All rights reserved.
Lab Protocol Title
Current Protocols in Protein Science
How to Cite
Kemp, G., Fliegel, L. and Young, H. S. 2014. Membrane Transport Piece by Piece: Production of Transmembrane Peptides for Structural and Functional Studies. Current Protocols in Protein Science. 75:29.8:29.8.1–29.8.28.
- Published Online: 3 FEB 2014
Membrane proteins are involved in all cellular processes from signaling cascades to nutrient uptake and waste disposal. Because of these essential functions, many membrane proteins are recognized as important, yet elusive, clinical targets. Recent advances in structural biology have answered many questions about how membrane proteins function, yet one of the major bottlenecks remains the ability to obtain sufficient quantities of pure and homogeneous protein. This is particularly true for human membrane proteins, where novel expression strategies and structural techniques are needed to better characterize their function and therapeutic potential. One way to approach this challenge is to determine the structure of smaller pieces of membrane proteins that can be assembled into models of the complete protein. This unit describes the rationale for working with single or multiple transmembrane segments and provides a description of strategies and methods to express and purify them for structural and functional studies using a maltose binding protein (MBP) fusion. The bulk of the unit outlines a detailed methodology and justification for producing these peptides under native-like conditions. Curr. Protoc. Protein Sci. 75:29.8.1-29.8.28 © 2014 by John Wiley & Sons, Inc.
- membrane proteins;
- hydrophobic peptides;
- bacterial expression;
- maltose binding protein;
- organic extraction;
- molecular structure