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Detection of the pH-dependent Activity of Escherichia coli Chaperone HdeB In Vitro and In Vivo
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JoVE Journal Bioquímica
Detection of the pH-dependent Activity of Escherichia coli Chaperone HdeB In Vitro and In Vivo

Detection of the pH-dependent Activity of Escherichia coli Chaperone HdeB In Vitro and In Vivo

DOI:
10.3791/54527-v

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08:32 min

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October 23, 2016

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, , ,
1Department of Molecular, Cellular, and Developmental Biology,University of Michigan, 2Howard Hughes Medical Institute,University of Michigan

Capítulos

  • 00:05Título
  • 01:16Chaperone Activity Assay Using Thermally Unfolding Malate Dehydrogenase (MDH)
  • 03:18Effect of HdeB Overexpression on E. coli Survival under Acid Stress
  • 05:14Results: Chaperone Activity Assay Using Thermally Unfolded MDH and the Effect of HdeB Overexpression on E. coli Survival under Acid Stress
  • 07:16Conclusion

Summary

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This study describes biophysical, biochemical and molecular techniques to characterize the chaperone activity of Escherichia coli HdeB under acidic pH conditions. These methods have been successfully applied for other acid-protective chaperones such as HdeA and can be modified to work for other chaperones and stress conditions.

Tags

Keywords: Escherichia ColiChaperone HdeBPH-dependent ActivityIn VitroIn VivoAcid Activated ChaperonesProtein UnfoldingProtein DegradationEnteric BacteriaStomachAcid Protective ChaperonesHDAMalate DehydrogenaseLight ScatteringFluorescent SpectrophotometerTemperature Control

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