UID:
almafu_9958131492202883
Umfang:
1 online resource (494 p.)
Ausgabe:
1st ed.
ISBN:
9786613020369
,
9781283020367
,
128302036X
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9780123813480
,
0123813484
Serie:
Methods in enzymology, v. 471
Inhalt:
Multicellular organisms must be able to adapt to cellular events to accommodate prevailing conditions. Sensory-response circuits operate by making use of a phosphorylation control mechanism known as the ""two-component system."" This volume, the third in a three-volume treatment edited by the same group of editors, includes a wide range of methods, including those dealing with the Sln-1 kinase pathway, triazole sensitivity in C. albicans, and histidine kinases in cyanobacteria circadian clock. * Includes time-tested core methods and new innovations applicable to any res
Anmerkung:
Description based upon print version of record.
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Front Cover; Methods in Enzymology Two-Component Signaling Systems, Part C; Copyright Page; Contents; Contributors; Chapter 1: Characterizing Cross-Talk In Vivo: Avoiding Pitfalls and Overinterpretation; Abstract; 1. Overview; 2. Sources of Cross-Talk; 3. Cross-Talk Suppression; 4. Transcriptional Reporters; 5. Response Regulator Localization; 6. Phosphatase Cross-Talk; 7. Signal Response in Cross-Talk Networks; 8. Concluding Remarks; Acknowledgments; References; Chapter 2: Inference of Direct Residue Contacts in Two-Component Signaling; Abstract; 1. Introduction; 2. Extraction Tools
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3. DCA: Direct Coupling AnalysisAppendix: Nonstandard Linear Algebra Functions; Acknowledgments; References; Chapter 3: Computational Modeling of Phosphotransfer Complexes in Two-Component Signaling; Abstract; 1. Introduction; 2. Methods; 3. Summary; Acknowledgments; References; Chapter 4: Kinetic Studies of the Yeast His-Asp Phosphorelay Signaling Pathway; Abstract; 1. Introduction; 2. Materials and Methods; 3. Conclusion; Acknowledgments; References; Chapter 5: Purification of MBP-EnvZ Fusion Proteins Using an Automated System; Abstract; 1. Introduction
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2. Comparison Between the Cytoplasmic Domains of E. coli and Typhi EnvZ Proteins3. Purification of the Recombinant MBP Proteins by FPLC; 4. SDS-PAGE Analysis of Recombinant Expressed MBP-EnvZc Protein; 5. Results; 6. Discussion; Acknowledgments; References; Chapter 6: Measurement of Response Regulator Autodephosphorylation Rates Spanning Six Orders of Magnitude; Abstract; 1. Overview of Response Regulator Autodephosphorylation; 2. Purification of Response Regulator Proteins; 3. General Considerations for Autodephosphorylation Assays; 4. Assay of Autodephosphorylation by Loss of 32P
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5. Assay of Autodephosphorylation by Fluorescence6. Assay of Autodephosphorylation by Pi Release; 7. Assay of Autodephosphorylation from Systems of Reactions; 8. Future Prospects; Acknowledgments; References; Chapter 7: Transmembrane Receptor Chimeras to Probe HAMP Domain Function; Abstract; 1. Introduction; 2. Design of Chimeras of HAMP Domains Fused to the AC Rv3645 Catalytic Domain; 3. Choice of Vector and Cloning Strategy; 4. Expression and Purification of the Chimeras; 5. AC Assay; 6. Example of Applications of the Method and Results; 7. Concluding Remarks; Acknowledgments; References
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Chapter 8: Light-Activated Bacterial LOV-Domain Histidine KinasesAbstract; 1. Introduction; 2. Description of Method; 3. Concluding Remarks and Future Perspectives; Acknowledgments; References; Chapter 9: Characterization of Bacteriophytochromes from Photosynthetic Bacteria; Abstract; 1. Introduction; 2. Cloning BphP in Expression Vector; 3. Overexpression and Purification of BphPs; 4. Autophosphorylation; 5. Phosphotransfer; 6. Gel Mobility Shift Assay and DNase I Footprint; 7. Gene Disruption; 8. Analysis of the Photosynthetic Phenotypes of the BphP Mutants; 9. Photochemical Measurements
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References
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English
Weitere Ausg.:
ISBN 9780123813473
Weitere Ausg.:
ISBN 0123813476
Sprache:
Englisch
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