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Membrane proteins. Engineering, purification and crystallization / edited by Arun K. Shukla.

Contributor(s): Material type: TextTextSeries: Methods in enzymology ; v. 557.Publisher: Waltham, MA : Academic Press, 2015Copyright date: ©2015Description: 1 online resource (xx, 624 pages) : illustrations (chiefly color)Content type:
  • text
Media type:
  • computer
Carrier type:
  • online resource
ISBN:
  • 9780128021934
  • 0128021934
Other title:
  • Engineering, purification and crystallization
Subject(s): Genre/Form: Additional physical formats: Print version:: Membrane proteins - Engineering, Purification and Crystallization.DDC classification:
  • 572/.696 23
LOC classification:
  • QP552.M44
Online resources:
Contents:
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.
Summary: 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 interested audience. Illustrations and case example images are included wherever appropriate to help the readers understand the schematics and general experimental outlines. Volume of Methods In EnzymologyContains a collection of a diverse array of topics in the area of membrane protein biology rangin.
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Includes bibliographical references and indexes.

Online resource; title from title details screen (ScienceDirect, viewed May 5, 2015).

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.

880-01 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.

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 interested audience. Illustrations and case example images are included wherever appropriate to help the readers understand the schematics and general experimental outlines. Volume of Methods In EnzymologyContains a collection of a diverse array of topics in the area of membrane protein biology rangin.

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