Small GTPases in disease. Pt. A / edited by William E. Balch, Channing J. Der, Alan Hall.

Format:

Book

Contributor:

Balch, William Edward, 1949-

Der, Channing J.

Hall, A. (Alan)

Series:

Methods in enzymology ; v. 438.

Methods in enzymology ; v. 438

Small GTPases in disease

Language:

English

Subjects (All):

Guanosine triphosphatase.

Enzymology.

Physical Description:

1 online resource (512 p.)

Place of Publication:

Amsterdam : Academic Press/Elsevier, 2008.

Language Note:

English

Summary:

The critically acclaimed laboratory standard for more than forty years, Methods in Enzymology is one of the most highly respected publications in the field of biochemistry. Since 1955, each volume has been eagerly awaited, frequently consulted, and praised by researchers and reviewers alike. Now with over 400 volumes (all of them still in print), the series contains much material still relevant today-truly an essential publication for researchers in all fields of life sciences. Methods in Enzymology is now available online at ScienceDirect - full-text online of volumes 1 onw

Contents:

Front Cover; Small GTPases in Disease, Part A; Copyright Page; Contents; Contributors; Volumes in Series; Chapter 1: Rab1b Silencing Using Small Interfering RNA for Analysis of Disease-Specific Function; 1. Introduction; 2. Materials for siRNA Silencing of Rab1b; 3. Procedure for Transfection of siRNA; 4. Titration of Optimal siRNA Concentration; 5. SDS-PAGE Analysis of Rab1 Knockdown; 6. SDS-PAGE Analysis of CFTR; 7. Discussion; References; Chapter 2: Rab8-Optineurin-Myosin VI: Analysis of Interactions and Functions in the Secretory Pathway; 1. Introduction

2. Identification of Myosin VI-Binding Partners3. Verification of Yeast Two-Hybrid Screen Results; 4. Mammalian Two-Hybrid Assay; 5. Protocol for Mammalian Two-Hybrid Assay; 6. SEAP Secretion Assay; 7. Measurement of SEAP Secretion in Fibroblasts from Myosin VI Knockout Mouse; 8. Measurement of SEAP Secretion in siRNA Knockdown Cells; 9. Protocol for SEAP Assay; Acknowledgments; References; Chapter 3: Characterization of Rab27a and JFC1 as Constituents of the Secretory Machinery of Prostate-Specific Antigen in Prostate Carcinoma Cells; 1. Introduction; 2. Methods; 3. Concluding Remarks

AcknowledgmentsReferences; Chapter 4: In Vitro Assays to Characterize Inhibitors of the Activation of Small G Proteins by Their Guanine Nucleotide Exchange Factors; 1. GEFs and Diseases: Inhibiting the Exchange Reaction by Small Molecules; 2. Initial Screen of Inhibitors of SMG Activation; 3. Determination of Inhibition Profile by Using Michaelis-Menten Formalism; 4. Complementary Assays to Refine.Noncompetitive and Uncompetitive Mechanisms; 5. Conclusion; Acknowledgments; References; Chapter 5: Analysis of Small GTPase Function in Trypanosomes; 1. Overview; 2. Culturing

3. Transfection and Cloning of T. brucei4. Vector Systems for Overexpression and RNA Interference; 5. Cloning and Manipulation of Trypanosome Small GTPase Sequences; 6. Trafficking Assays to Monitor Exocytosis and Endocytosis; Acknowledgments; References; Chapter 6: Use of Dynasore, the Small Molecule Inhibitor of Dynamin, in the Regulation of Endocytosis; 1. Introduction; 2. Dynamin; 3. Dynamin and the Actin Cytoskeleton; 4. The ""Chemical Genetics"" Discovery Approach; 5. Why Do We Need Interfering Small Molecules?; 6. Synthesis of Dynasore; 7. Storage Conditions for Dynasore

8. Expression, Purification, and Storage of Dynamin9. Buffers and Reagents; Acknowledgment; References; Chapter 7: Identification and Verification of Sro7p as an Effector of the Sec4p Rab GTPase; 1. Introduction; 2. Experimental Approach to the Identification of Sec4p Effectors; 3. Purification of GST-Sec4, GST-Ypt1, and GST, and Nucleotide Exchange; 4. Preparation of the Yeast Extract for Sec4p Affinity Chromatography; 5. Sec4p Affinity Chromatography; 6. Confirmation of Sro7p as a Sec4p Effector; 7. Co-Immunoprecipitation of Sro7p with Sec4p; References

Chapter 8: Characterization of Rab18, a Lipid Droplet-Associated Small GTPase

Notes:

Description based upon print version of record.

Includes bibliographical references and indexes.

ISBN:

9786611308711

9781281308719

1281308714

9780080877600

0080877605

OCLC:

808681976