John Graham, Joan Higgins

Membrane Analysis (eBook, ePUB)

• Format: ePub

Produktbeschreibung

This book provides a comprehensive review of laboratory methods for membrane study, with an emphasis on isolating membranes, analysing their composition and architecture, and investigating membrane function.

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Produktdetails

• Verlag: Taylor & Francis
• Seitenzahl: 224
• Erscheinungstermin: 13. August 2020
• Englisch
• ISBN-13: 9781000141023
• Artikelnr.: 59870122

Autorenporträt

John Graham Research Consultancy, Peter Jost Enterprise Centre, Liverpool John Moores University, Byrom Street, Liverpool L3 3AF, UK. The University of Sheffield, Department of Molecular Biology and Biotechnology, P.O. Box 594, Firth Court, Western Bank, Sheffield Sl0 2TN, UK.

Inhaltsangabe

Abbreviations
Preface
1. General membrane composition and structure
Lipids
General lipid composition
Phospholipids
Isoprenoids
Lipid architecture
Proteins
Peripheral proteins
Integral proteins
Lipid
anchored proteins
Protein architecture in the plane of the membrane
Carbohydrates
Glycolipids
Glycoproteins
Proteoglycans
2. The membranes of prokaryotes and eukaryotes
General features
Membranes of prokaryotes
Membranes of eukaryotes
The surface (plasma) membrane
Plasma membrane domains
The cytoskeleton and glycocalyx
Mitochondria and chloroplasts
Peroxisomes (microbodies)
Lysosomes
Nuclei
Endocytic compartment
The secretory compartment
References
3. Preparation of subcellular membranes
Homogenization
Liquid shear
Gaseous shear
Mechanical shear
Other methods
Homogenization of soft mammalian tissue
Homogenization of tough tissues
Homogenization of cultured mammalian cells
Plant tissue
Yeast
Bacteria
Fractionation of subcellular organelles
Differential centrifugation
Density gradient centrifugation
Density perturbation
Fractionation by immunoaffinity
Fractionation by continuous flow electrophoresis
Fractionation in aqueous two
phase systems
Identification of fractionated membranes
Morphological
Chemical
Enzyme markers
Immunological markers
Expression of data
Some specimen separations of organelles and membranes
Nuclei
Golgi membranes
Mitochondria
Resolving a light mitochondrial fraction
Plasma membrane
Membrane vesicles
References
4. Molecular composition
Chemical analysis of proteins
Quantitative assay
Separation of membrane proteins by sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS
PAGE)
Western blotting (immunoblotting)
Resolution of proteins by isoelectric focusing and twodimensional (2D) electrophoresis
Chemical analysis of lipids
Lipid extraction
Analysis of phospholipids
Separation and analysis of phospholipids by HPLC
Analysis of phospholipid fatty acid
Determination of cholesterol
Separation and analysis of glycolipids
Glycoprotein oligosaccharide analysis
Glycoprotein analysis
Preparation of a crude glycoprotein fraction
Glycoprotein fractionation
Chemical assays for carbohydrates
Isolation of the oligosaccharide chains
Oligosaccharide sequence analysis
References
5. Investigating the topology of membrane proteins
Demonstration of the orientation of membrane vesicles
Identification and separation of peripheral and integral proteins
Triton X114 as a reagent for separation of integral and peripheral proteins
Identification of proteins anchored by phosphatidylinositol
Identification of lipid
modified proteins
Probing the topography of integral membrane proteins
General experimental criteria
Probes for integral membrane proteins
Hydropathy plots for prediction of membrane protein topography
Structural analysis of membrane proteins
References
6. Investigation of membrane lipids
Determination of the organization of membrane phospholipids
Model membranes
Multilamellar vesicles (MLVs)
Small unilamellar vesicles (SUVs)
Large unilamellar vesicles (LUVs)
Black lipid membranes
Reconstituted membranes
Determination of the transverse distribution of membrane phospholipids
Chemical probes
Phospholipases
Phospholipid transfer proteins
Assay based on prothrombinase
Determination of the transverse distribution of membrane cholesterol
Studies of lipid transport
Investigation of the transmembrane movement of phospholipids
Transport of phospholipids and cholesterol between intracellular membranes
Genetic approach to investigate membrane phospholipids
References
7. Protein targetting and membrane biogenesis
Genetic approaches to studies of protein targetting
Biochemical approaches to studies of protein targetting
Steps in protein targetting
Nuclear targetting
Targetting to peroxisomes
Targetting to mitochondria
Targetting to the ER
Transport across the ER membrane
Retention in the ER lumen
Transfer to the Golgi apparatus
Targetting to lysosomes
Transit of mannose
6
phosphatase receptors to and from the plasma membrane and Golgi membranes
Establishing the topography and location of the membrane proteins of the ER, Golgi and plasma membrane
Covalently bound lipids in protein targetting
Targetting in prokaryotes
Overview
References
8. Endocytosis and vesicular trafficking
Receptors and endocytosis
Receptor
mediated endocytosis
Membrane receptors
Investigation of events in endocytosis
Electron microscopy
Biochemical studies
Isolation of transport vesicles and investigation of the factors involved in vesicular transport
Reconstitution of vesicular transport
Semi
intact cell systems
Transfer vesicles
Transfer from the plasma membrane to the endosomes
Transfer from the Golgi to lysosomes
Golgi to endoplasmic reticulum (retrograde transport)
Endoplasmic reticulum to Golgi (anterograde transport)
Recognition in vesicular transfer
References
Index.