Current strategies for protein production and purification enabling membrane protein structural biology.
TLDR
This review focuses on both the advances and diversity of protein production and purification methods that have allowed this growth in structural knowledge of membrane proteins through X-ray crystallography, nuclear magnetic resonance spectroscopy, and cryo-electron microscopy (cryo-EM).Abstract:
Membrane proteins are still heavily under-represented in the protein data bank (PDB), owing to multiple bottlenecks. The typical low abundance of membrane proteins in their natural hosts makes it necessary to overexpress these proteins either in heterologous systems or through in vitro translation/cell-free expression. Heterologous expression of proteins, in turn, leads to multiple obstacles, owing to the unpredictability of compatibility of the target protein for expression in a given host. The highly hydrophobic and (or) amphipathic nature of membrane proteins also leads to challenges in producing a homogeneous, stable, and pure sample for structural studies. Circumventing these hurdles has become possible through the introduction of novel protein production protocols; efficient protein isolation and sample preparation methods; and, improvement in hardware and software for structural characterization. Combined, these advances have made the past 10-15 years very exciting and eventful for the field of membrane protein structural biology, with an exponential growth in the number of solved membrane protein structures. In this review, we focus on both the advances and diversity of protein production and purification methods that have allowed this growth in structural knowledge of membrane proteins through X-ray crystallography, nuclear magnetic resonance (NMR) spectroscopy, and cryo-electron microscopy (cryo-EM).read more
Citations
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A systematic approach to quantitative Western blot analysis.
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Folding and Misfolding of Human Membrane Proteins in Health and Disease: From Single Molecules to Cellular Proteostasis
Justin T. Marinko,Hui Huang,Wesley D. Penn,John A. Capra,Jonathan P. Schlebach,Charles R. Sanders +5 more
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Plant lipids: Key players of plasma membrane organization and function.
Adiilah Mamode Cassim,Paul Gouguet,Julien Gronnier,Nelson Laurent,Véronique Germain,Magali S. Grison,Yohann Boutté,Patricia Gerbeau-Pissot,Françoise Simon-Plas,Sébastien Mongrand +9 more
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Apelinergic System Structure and Function
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Diblock copolymers enhance folding of a mechanosensitive membrane protein during cell-free expression
TL;DR: It is suggested that global membrane physical properties, specifically available membrane surface area and the membrane area expansion modulus, significantly influence the folding and yield of a membrane protein.
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