Chaperonin protocols / edited by Christine Schneider.

Format:

Book

Contributor:

Schneider, Christine, 1942-

Series:

Methods in molecular biology (Clifton, N.J.) ; v. 140.

Methods in molecular biology ; v. 140

Language:

English

Subjects (All):

Molecular chaperones--Purification--Laboratory manuals.

Molecular chaperones.

Chaperonins--analysis.

Chaperonins--isolation & purification.

Medical Subjects:

Chaperonins--analysis.

Chaperonins--isolation & purification.

Genre:

Laboratory manuals.

Physical Description:

x, 212 pages : illustrations ; 23 cm.

Place of Publication:

Totowa, N.J. : Humana Press, [2000]

Summary:

In Chaperonin Protocols, Christine Schneider has assembled a unique collection of readily reproducible protocols for the study of chaperonins, intracellular proteins critical to many biological processes. Written by experienced investigators who have successfully honed their methods to a fineness, the protocols focus on the purification of chaperonins from different species along with their corresponding cofactors, and on chaperonin activity assays for in vivo as well as in vitro work. Many activity assays are given for GroEL, which can also be applied to mitochrondrial Hsp60. There are also assays for the eukaryotic chaperonin TRiC and handy methods--for example, one for preparing labeled probes--that can be used for various purposes and prove helpful in numerous different procedures.

Critically important to a greater understanding of such disorders as cystic fibrosis, Alzheimer's disease, and BSE, Chaperonin Protocols offers both novice and experienced investigators fast access to today's best and most productive chaperonin methods, all explained in step-by-step detail to ensure robust and reproducible results.

Contents:

1 Purification of Archaeal Chaperonin from Sulfolobus shibatae / Elsie Quaite-Randall, Andrzej Joachimiak 1

2 Purification of Hsp60 from Thermus thermophilus / Elsie Quaite-Randall, Andrzej Joachimiak 15

3 Purification of GroEL from an Overproducing E. coli Strain / Elsie Quaite-Randall, Andrzej Joachimiak 29

4 Purification of GroES from an Overproducing E. coli Strain / Elsie Quaite-Randall, Andrzej Joachimiak 41

5 Purification of the Gp31 Co-chaperonin from Bacteriophage T4 / Saskia M. van der Vies 51

6 Removing Trace Fluorescent Contaminants from GroEL Preparations / Frank Weber 63

7 Assembly and Disassembly of GroEL and GroES Complexes / Jorg Martin 65

8 GroEL/GroES Interaction Assayed by Protease Protection / Jorg Martin 71

9 Determination of Chaperonin Activity In Vivo / Saskia M. van der Vies, Peter A. Lund 75

10 Interaction of Nonnative Polypeptide Substrates with the Escherichia coli Chaperonin GroEL / Wanda K. Hartmann, Edward Eisenstein 97

11 Prevention of Rhodanese Aggregation by the Chaperonin GroEL / Frank Weber, Manajit Hayer-Hartl 111

12 Refolding of Bovine Mitochondrial Rhodanese by Chaperonins GroEL and GroES / Frank Weber, Manajit Hayer-Hartl 117

13 Assay of Malate Dehydrogenase: A Substrate for the E. coli Chaperonins GroEL and GroES / Manajit Hayer-Hartl 127

14 Assay of Chaperonin-Assisted Refolding of Citrate Synthase / N. Kalaya Steede, Stacy L. Temkin, Samuel J. Landry 133

15 Purification of Yeast Mitochondrial Hsp60 / Yves Dubaquie, Renate Looser, Sabine Rospert 139

16 Preparation of Recombinant Human Hsp 10 / N. Kalaya Steede, Jesse J. Guidry, Samuel J. Landry 145

17 Purification of the Cytosolic Chaperonin TRiC from Bovine Testis / Raul G. Ferreyra, Judith Frydman 153

18 Monitoring Actin Folding: Purification Protocols for Labeled Proteins and Binding to DNase I-Sepharose Beads / Vanitha Thulasiraman, Raul G. Ferreyra, Judith Frydman 161

19 Folding Assays: Assessing the Native Conformation of Proteins / Vanitha Thulasiraman, Raul G. Ferreyra, Judith Frydman 169

20 Purification of Prefoldin / Sally A. Lewis, Nicholas J. Cowan 179

21 Purification of GimC from Saccharomyces cerevisiae / Katja Siegers, Elmar Schiebel 185

22 Analysis of Eukaryotic Molecular Chaperone Complexes Involved in Actin Folding / Michel R. Leroux 195.

Notes:

Includes bibliographical references and index.

ISBN:

0896037398

OCLC:

42049240