Membrane proteins. Engineering, purification and crystallization /

• Άλλοι συγγραφείς: Shukla, Arun K. (Επιμελητής έκδοσης)
• Μορφή: Ηλ. βιβλίο
• Γλώσσα: English
• Έκδοση: Waltham, MA : Academic Press, 2015.
• Σειρά: Methods in enzymology ; v. 557.
• Θέματα: Membrane proteins.; SCIENCE > Life Sciences > Biochemistry.; Membrane Proteins.; Electronic books.

Membrane Proteins - Engineering, Purification and Crystallization, a volume of Methods In Enzymology, encompasses chapters from the leading experts in the area of membrane protein biology. The chapters provide a brief overview of the topics covered and also outline step-by-step protocol for the inte...

Πίνακας περιεχομένων:

• Front Cover; Membrane Proteins-Engineering, Purification and Crystallization; Copyright; Contents; Contributors; Preface; Section I: Membrane Protein Engineering, Solubilization and Purification; Chapter One: Multicolor Fluorescence-Based Screening Toward Structural Analysis of Multiprotein Membrane Complexes; 1. Introduction; 1.1. Screening approaches for membrane proteins and membrane protein complexes; 1.2. Multicolor fluorescence-based screening of multiprotein membrane complexes.
• 1.3. The major histocompatibility complex class I peptide-loading complex: A challenge for structural biology of membrane ... 2. Production of Fluorescently Labeled TAP Complexes in P. pastoris; 2.1. Construct design; 2.2. Transformation; 2.3. Colony screening; 2.4. Large-scale expression; 2.5. Orthogonal purification; 3. Production of Fluorescently Labeled TAP Complexes in Mammalian Cells; 3.1. Construct design; 3.2. Transfection; 3.3. Orthogonal purification; 4. Multicolor Fluorescence-Based Screening Approaches; 4.1. MC-FSEC analysis; 4.2. In-gel fluorescence and immunoblotting.
• 4.3. Monitoring peptide binding to TAP4.4. Measurement of MHC I surface presentation by flow cytometry; 5. Summary; Acknowledgments; References; Chapter Two: A Novel Screening Approach for Optimal and Functional Fusion of T4 Lysozyme in GPCRs; 1. Introduction; 2. Overall Strategy; 3. Plasmids, Yeast Strains, and Media; 3.1. Yeast strains; 3.2. Plasmids; 3.3. Media; 4. Library Construction; 4.1. Generation of T4L-containing cassettes with variable lengths of flanking regions; 4.2. Insertion of cassettes containing the T4L gene into the replacement region of the receptor.
• 2. MP Stability in Membrane Architecture3. Conventional Detergents; 3.1. Detergent classification; 3.2. Use of conventional detergents for MP study; 3.3. Limitation of conventional detergents; 4. Novel Amphipathic Systems; 4.1. Detergent properties for MP structural study; 4.2. Small amphipathic compounds; 4.2.1. Conventional detergent variants; 4.2.2. Hemifluorinated surfactants; 4.2.3. Tripod amphiphiles; 4.2.4. Facial amphiphiles; 4.2.5. Rigid hydrophobic group-bearing amphiphiles; 4.2.6. Neopentyl glycol (NG) class amphiphiles; 4.3. Peptide-based amphiphiles.