Structural Biology of Membrane Proteins (eBook)

Part One: Expression and Purification of Membrane Proteins; Chapter 1: Refolding of G-protein-coupled receptors; 1: Introduction; 2: Refolding of membrane proteins; 3: In vitro protein refolding; 4: GPCR in vitro refolding; 5: Conclusion; 6: References; Chapter 2: Expression of genes encoding eukaryotic membrane proteins in mammalian cells; 1: Introduction; 2: Mammalian cell hosts and gene expression vectors; 3: Delivery and maintenance of expression vectors in mammalian host cells; 4: Use of HEK293S stable cell lines for high-level expression of eukaryotic membrane proteins; 5: Large-scale growth of HEK293S stable cell lines in suspension culture using a bioreactor and inducible gene expression; 6: Preparation of eukaryotic membrane proteins containing simple N-glycans; 7: Outlook for use of HEK293S tetracycline-inducible cell lines for large-scale preparation of other eukaryotic membrane proteins; Chapter 3: Expression of recombinant G-protein-coupled receptors for structural biology; 1: Introduction; 2: Expression of recombinant GPCRs; 3: Conclusions; 4: References; Chapter 4: The purification of G-protein-coupled receptors for crystallization; 1: Introduction; 2: Heterogeneity of overexpressed receptors; 3: Membrane fractionation, solubilization and detergent selection; 4: Purification; 5: Final quality control, monitoring protein stability, aggregational state, lipid and bound detergent; 6: Conclusions; Chapter 5: An introduction to detergents and their use in membrane protein studies; 1: Introduction; 2: Physical properties of detergents used in membrane protein studies; 3: Extraction and purification procedure using common detergents; 4: Use of detergents in membrane protein crystallization; 5: Conclusion; 6: References; Part Two: Methods for Structural Characterization of Membrane Proteins; Chapter 6: Solution NMR approaches to the structure and dynamics of integral membrane proteins; 1: Introduction; 2: Protein production and optimization for NMR studies; 3: NMR methodology for the study of integral membrane proteins; 4: Solution NMR structures of helical integral membrane proteins; 5: Solution NMR Structures of ?-barrel membrane proteins; 6: Solution NMR characterization of membrane protein dynamics; 7: Future directions; 8: References; Chapter 7: Membrane proteins studied by solid-state NMR; 1: Introduction; 2: Sample preparation and methodology; 3: Applications; 4: Conclusions; 5: References; Chapter 8: Assessing structure and dynamics of native membrane proteins; 1: Introduction; 2: Assembly of 2D crystals; 3: Electron microscopy; 4: Atomic force microscopy; 5: Conclusion and perspectives; Chapter 9: State-of-the-art methods in electron microscopy, including single particle analysis; 1: Introduction; 2: Sample Preparation; 3: Low Dose Microscopy; 4: Applications of CryoEM; 5: Examples; 6: Conclusion; Chapter 10: Atomic resolution structures of integral membrane proteins using cubic lipid phase crystallization; 1: Introduction; 2: Membrane protein crystals & crystallization; 3: Advantages of structures in a native setting at high resolution; 4: Conclusions; 5: References; Part Three: New Membrane Protein Structures; Chapter 11: Aquaporins: Integral membrane channel proteins; 1: Introduction; 2: Aquaporin channel proton exclusion barrier; 3: Selectivity in the aquaporin family; 4: Permeation by substances other than water and glycerol; 5: Aquaporin monomer associations and their functional implications; 6: References; Chapter 12: Gas channels for ammonia; 1: Introduction; 2: The structure of Ammonia Channel; 3: Reconstituted into liposomes AmtB acts as a channel that conducts NH3; 4: The Mechanism of conduction; 5: The Rh proteins; 6: Comparison with AQPs; 7: Comparison with K+ channel; 8: Acknowledgement; 9: References; Chapter 13: Channels in the outer membrane of Mycobacter; 1: Introduction; 2: Structure determination; 3: Structure description; 4: The outer membrane; 5: Conclusion; 6: References; Chapter 14: The structure of the SecY protein translocation channel; 1: Abstract; 2: Introduction; 3: Structure determination of the SecY complex by electron cryo-microscopy; 4: Determination of the X-ray crystal structure of the SecY complex; 5: Description of the structure of the SecY complex; 6: Post-translational translocation in bacteria; 7: Conclusions and outlook; 8: References; Chapter 15: Structure and function of the translocator domain of bacterial autotransporters; 1: Introduction; 2: The NalP autotransporter; 3: The translocator domain of autotransporters; 4: Purification and in vitro folding of the NalP translocator domain; 5: The structure of the NalP translocator domain; 6: Comparison of the NalP translocator domain to other translocator domains and to TolC; 7: The autotransporter secretion mechanism; Chapter 16: X-ray crystallographic structures of sarcoplasmic reticulum Ca2+-ATPase at the atomic level; 1: Introduction; 2: The transport scheme and thermodynamics of Ca2+ transport; 3: Overall structure of Ca2+-ATPase; 4: Transport models; 5: Initialization of the cycle - phosphorylation and calcium ion occlusion; 6: The dephosphorylation step and proton counter transport; 7: Getting Ca2+ in and out of the membrane; 8: Compact versus open conformations of SERCA; 9: Conclusions; Chapter 17: Comparison of the multidrug transporter EmrE structures determined by electron cryo-microscopy and X-ray crystallography; 1: Introduction; 2: The oligomeric state of EmrE; 3: Transport activity of EmrE; 4: Structure of EmrE determined by electron cryomicroscopy; 5: Comparison of the EmrE structure determined by electron crystallography with a 3.8 + resolution structure determined by X-ray crystallography; 6: Conclusion; 7: References; Chapter 18: Structure of photosystems I and II; 1: Introduction to oxygenic photosynthesis; 2: Photosystem ii; 3: Photosystem I; 4: Conclusion and outlook; Chapter 19: Glutamate receptor ion channels - Structural insights into molecular mechanism; 1: Introduction; 2: Studies of the Ligand-binding Domain; 3: The Functional Architecture of a Glutamate Receptor Ion Channel; 4: A Working Model of AMPA Receptor Function; 5: Open Questions; 6: References; Chapter 20: The mitochondrial ADP/ATP carrier; 1: Introduction; 2: Mitochondrial carriers and ADP/ATP carrier; 3: Crystallization; 4: Diffraction, phasing and model building; 5: Structure analysis; 6: Functional implications; 7: Future developments and conclusions;