Library Science

A collection of key techniques for the study of receptors and transport proteins. The book provides examples of how different membrane proteins can be overexpressed in both prokaryotic and eukaryotic expression systems, how natural and overexpressed proteins can be solubilized from their host membranes, and how the solubilized protein can be purified in active form. Each protocols contains step-by-step instructions to ensure success, troubleshooting advice, lists of reagents, and tips on avoiding pitfalls. Ed. B.S. Selinsky

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Table of Contents

1. Expression and Purification of the Amphipathic Form of Rabbit Cytochrome b5 in Escherichia coli (pp. 3-10)
   Waskell, Lucy
2. Dihydroorotate Dehydrogenase of Escherichia coli (pp. 11-22)
   Jensen, Kaj Frank; Larsen, Sine
3. A General Approach for Heterologous Membrane Protein Expression in Escherichia coli: The Uncoupling Protein, UCP1, as an Example (pp. 23-36)
   Shaw, Alison Z.; Miroux, Bruno
4. Expression of Membrane-Bound Iron-Sulfur Proteins (pp. 37-48)
   Schneider, Dirk; Schmidt, Christian L.; Seidler, Andreas
5. Heterologous Expression of Human Scavenger Receptor Class B Type I (SR-BI) in Pichia pastoris (pp. 49-64)
   Han, Chang-Hoon; Werder, Moritz; von Gustedt, Wilk; Hauser, Helmut
6. Expression of Oligomeric Amiloride-Sensitive Epithelial Sodium Channel in Sf9 Insect Cells (pp. 65-72)
   Rao, U. Subrahmanyeswara
7. Functional Expression of His-Tagged Rhodopsin in Sf9 Insect Cells (pp. 73-86)
   Bosman, Giel J.C.G.M.; van Oostrum, Jenny; Breikers, Githa; Bovee-Geurts, Petra H.M.; Klaassen, Corne H.W.; DeGrip, Willem J.
8. Preparation of Glycerol Facilitator for Protein Structure and Folding Studies in Solution (pp. 89-102)
   Manley, Darren; ONeil, Joe D.
9. Solubilization of Chemokine Receptors from Cell Membranes (pp. 103-110)
   Staudinger, Robert; Bandres, Juan C.
10. A Systematic Approach for the Solubilization of the Integral Membrane Protein Lysophospholipid: Acyl-Coa Acyltransferase (LAT) (pp. 111-128)
    Kerkhoff, Claus; Kaever, Volkhard
11. Purification of Omp50, a Minor Porin of Campylobacter jejuni (pp. 131-138)
    Bolla, Jean Michel
12. Purification of Porins from Mycobacterium smegmatis (pp. 139-150)
    Heinz, Christian; Roth, Eva; Niederweis, Michael
13. Isolation of the Melanocortin 5 Receptor: From cDNA Sequence to Isolating an Integral Membrane Protein (pp. 151-162)
    Clarke, Benjamin L.
14. Purification of Mammalian Serine Palmitoyltransferase, a Hetero-Subunit Enzyme for Sphingolipid Biosynthesis, by Affinity-Peptide Chromatography (pp. 163-174)
    Hanada, Kentaro; Nishijima, Masahiro
15. Purification of Phosphatidylglycerophosphate Synthase from Cultured Mammalian Cells (pp. 175-186)
    Kawasaki, Kiyoshi; Nishijima, Masahiro
16. Purification of Pancreastatin Receptor from Rat Liver Membranes (pp. 187-194)
    Sanchez-Margalet, Victor; Santos-Alvarez, Jose; Diaz-Troya, Sandra
17. Purification, Reconstitution, and Functional Characterization of Zinc Transporter from Rat Renal Brush Border Membranes (pp. 195-222)
    Prasad, Rajendra
18. Isolation of Lipid Raft-Associated Proteolipids (pp. 223-230)
    Millan, Jaime; de Marco, Maria C.; Qaidi, Mohammed; Batista, Alicia; Martin-Belmonte, Fernando; Alonso, Miguel A.
19. Purification of Membrane-Bound Catechol-O-Methyltransferase (pp. 231-238)
    Bonifacio, Maria Joao; Soares-da-Silva, Patricio
20. Purification and Characterization of Transporter Proteins from Human Erythrocyte Membrane (pp. 239-256)
    Wang, Da-Neng; Lemieux, M. Joanne; Boulter, Jonathan M.
21. Purification of the Human Erythrocyte PS-Stimulated Mg2+-ATPase: A Putative PS Flippase (pp. 257-270)
    Lyles, Jill V.; Cornely-Moss, Kathleen; Smith, Christine M.; Daleke, David L.
22. Reconstitution and Assay of Biogenic Membrane-Derived Phospholipid Flippase Activity in Proteoliposomes (pp. 271-280)
    Gummadi, Sathyanarayana N.; Hrafnsdottir, Sigrun; Walent, Jane; E.Watkins, William; Menon, Anant K.
23. Expression, Purification, and Reconstitution of Rat Liver Carnitine Palmitoyltransferase I (pp. 281-302)
    Brown, Nicholas F.
24. Crystallization in Lipidic Cubic Phases: A Case Study with Bacteriorhodopsin (pp. 305-316)
    Gordeliy, Valentin I.; Schlesinger, Ramona; Efremov, Rouslan; Buldt, Georg; Heberle, Joachim
25. Optical Biosensor Assay Using Retroviral Receptor Pseudotypes (pp. 317-328)
    Rucker, Joseph

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