Quantification of Bacterial Histidine Kinase Autophosphorylation Using a Nitrocellulose Binding Assay

Jonathan Fischer; Roger A. Johnson; Elizabeth Boon

doi:10.3791/55129

Journal

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生物化学

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使用硝酸纤维素结合分析细菌组氨酸激酶自身磷酸化的定量分析

Quantification of Bacterial Histidine Kinase Autophosphorylation Using a Nitrocellulose Binding Assay

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生物化学

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JoVE 杂志生物化学

Quantification of Bacterial Histidine Kinase Autophosphorylation Using a Nitrocellulose Binding Assay

使用硝酸纤维素结合分析细菌组氨酸激酶自身磷酸化的定量分析

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8,490 Views

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09:17 min

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January 11, 2017

DOI:

10.3791/55129-v

DOI:

10.3791/55129-v

•

09:17 min

January 11, 2017

•

1 Views

Jonathan Fischer¹, Roger A. Johnson², Elizabeth Boon¹

¹Department of Chemistry,Stony Brook University, ²Department of Physiology and Biophysics,Stony Brook University

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Summary

We report and demonstrate an optimized nitrocellulose binding assay that can be used to quantify autophosphorylation of purified bacterial histidine kinases. Our method has several advantages over traditional SDS-PAGE based techniques, providing a valuable alternative for characterizing these important proteins.