Production of membrane proteins : strategies for expression and isolation /: strategies for expression and isolation. (2011)

• Record Type: Book
• Further Information: Note: Edited by Anne Skaja Robinson.
• Other Names: Robinson, Anne Skaja, 1966-
• Contents: Preface; INTRODUCTION; Expression; Solubilization and Structural Methods; Abbreviations; ; PART I: Expression Systems; ; BACTERIAL SYSTEMS; Introduction; Understanding the Problem; Vector/Promoter Types; T7 Expression System; Tunable T7 Expression Systems; Other Useful Membrane Protein Expression Strains; Clone Stability; Media Types; Fusion Partners/Membrane Targeting Peptides; Chaperone Overexpression; Cautionary Notes Related to Chaperone Overexpression; Emerging Role of Quality Control Proteases; Tag Selection; Potential Expression Yield; Strategies to Overcome Protein Instability; MEMBRANE PROTEIN EXPRESSION IN SACCHAROMYCES CEREVISIAE; Introduction; Getting Started; Special Considerations; Case Studies; Conclusions; EXPRESSION SYSTEMS: PICHIA PASTORIS; Introduction; A (Brief) Summary on the (Long) History of P. pastoris; Introducing P. pastoris as a Biotechnological Tool: Its (Extended) Strengths and (Limited) Weaknesses; Basics of the P. pastoris Expression System; Successful Large-Scale Expression of Membrane Proteins Using P. pastoris; Guidelines for Optimizing Membrane Protein Expression in P. pastoris Using GPCRs as Models; Conclusions and Future Directions; HETEROLOGOUS PRODUCTION OF ACTIVE MAMMALIAN G-PROTEIN-COUPLED RECEPTORS USING BACULOVIRUS-INFECTED INSECT CELLS; Introduction; Experimental; Conclusions and Future Perspectives; MEMBRANE PROTEIN EXPRESSION IN MAMMALIAN CELLS; Introduction; Mammalian Systems; Case Studies; Conclusions; MEMBRANE PROTEIN PRODUCTING USING PHOTOSYNTHETIC BACTERIA: A PRACTICAL GUIDE; Introduction; Preparation of Expression Constructs; Transfer of Plasmid DNA to Rhodobacter via Conjugal Mating; Small-Scale Screening for Expression and Localization of Target Protein in Rhodobacter; Large-Scale Culture; Detergent Solubilization and Chromatographic Purification of Expressed Membrane Proteins; Protein Identification and Assessment of Purity; Preparations of Specialized Rhodobacter Membranes; Appendix: Media and Buffer Formulations; ; PART II: Protein-Specific Considerations; ; PERIPHERAL MEMBRANE PROTEIN PRODUCTION FOR STRUCTURAL AND FUNCTIONAL STUDIES; Introduction; Case Studies of Peripheral Membrane Proteins; Conclusions; EXPRESSION OF G-PROTEIN-COUPLED RECEPTORS; Introduction; Bacterial Expression of GPCRs; Expression of GPCrs in Inclusion Bodies and Refolding; Expression of GPCrs in Yeast; Expression of GPCrs in Insect Cells; Expression of GPCrs in Mammalian Cell Lines; Expression of GPCrs in Retina Rod Cells; Expression of GPCrs in a Cell-Free System; Stabilization of GPCrs during Solubilization and Purification; Conclusions; STRUCTURAL BIOLOGY OF MEMBRANE PROTEINS; Introduction; Folding and Structural Analysis of Membrane Proteins; Test Cases; ; PART III: Emerging Methods and Approaches; ; ENGINEERING INTEGRAL MEMBRANE PROTEINS FOR EXPRESSION AND STABILITY; Introduction; Engineering Higher Expression; Engineering Higher Stability; Conclusions; EXPRESSION AND PURIFICATION OF G-PROTEIN-COUPLED RECEPTORS FOR NUCLEAR MAGNETIC RESONANCE STRUCTURAL STUDIES; Introduction: G-Protein-Coupled Receptor Superfamily; CXCR1; GPCR Structures; NMR Studies of GPCRs; Expression Systems; Cloning of CXCR1 into pGEX2a; Expression of CXCR1; Purification; Refolding and Reconstitution; Binding and Activity Measurements; NMR Spectra; SOLUBILIZATION, PURIFICATION, AND CHARACTERIZATION OF INTEGRAL MEMBRANE PROTEINS; Introduction; Solubilization of IMPs; IMP Purification; Characterization of Solubilized IMPs; STABILIZING MEMBRANE PROTEINS IN DETERGENT AND LIPID SYSTEMS; Introduction; Choice of Detergent: Solubilization versus Stability; Mitigating Protein Denaturation; Making or Selecting a Stable Protein; Conclusions; RAPID OPTIMIZATION OF MEMBRANE PROTEIN PRODUCTION USING GREEN FLUORESCENT PROTEIN-FUSIONS AND LEMO21(DE3); Introduction; Main Protocol; Materials; Expression and Isolation of GFP-His8; Conclusions; ;
• Publisher Details: Place of publication not identified : Wiley-VCH
• Publication Date: 2011
• Extent: 1 online resource (442 pages)
• Subjects: 572.696; Membrane proteins
• Languages: English
• ISBNs: 9783527634538
• Access Rights: Legal Deposit; Only available on premises controlled by the deposit library and to one user at any one time; The Legal Deposit Libraries (Non-Print Works) Regulations (UK).
• Access Usage: Restricted: Printing from this resource is governed by The Legal Deposit Libraries (Non-Print Works) Regulations (UK) and UK copyright law currently in force.