Exploration of the Effects of γ‑Phosphate-Modified ATP Analogues on Histidine Kinase Autophosphorylation

While two-component systems (TCSs), composed of a sensor histidine kinase (HK) and a response regulator, are the main signaling pathways in bacteria, global TCS activity remains poorly described. Here, we report the kinetic parameters of the HK autophosphorylation reaction using previously uncharact...

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Bibliographic Details
Published in:	Biochemistry (Easton) Vol. 57; no. 29; pp. 4368 - 4373
Main Authors:	Chase, Olivia M, Espinasse, Adeline, Wilke, Kaelyn E, Carlson, Erin E
Format:	Journal Article
Language:	English
Published:	United States American Chemical Society 24-07-2018
Subjects:	Adenosine Triphosphate - analogs & derivatives Adenosine Triphosphate - metabolism Bacterial Proteins - chemistry Bacterial Proteins - metabolism Histidine - chemistry Histidine - metabolism Histidine Kinase - chemistry Histidine Kinase - metabolism Kinetics Molecular Probes - chemistry Molecular Probes - metabolism Phosphorylation Thermotoga maritima - chemistry Thermotoga maritima - enzymology Thermotoga maritima - metabolism
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Summary:	While two-component systems (TCSs), composed of a sensor histidine kinase (HK) and a response regulator, are the main signaling pathways in bacteria, global TCS activity remains poorly described. Here, we report the kinetic parameters of the HK autophosphorylation reaction using previously uncharacterized γ-phosphate-modified ATP analogues to further elucidate their utility as activity-based probes for global TCS analysis. Given the increased stability of thiophosphorylated histidine in comparison to that of the native phosphoryl modification, which is attributed to the decreased electrophilicity of this moiety, we anticipated that ATPγS may be turned over much more slowly by the HKs. Surprisingly, we found this not to be the case, with the turnover numbers decreasing <1 order of magnitude. Instead, we found that alkylation of the thiophosphate had a much more dramatic effect on turnover and, in one case, the binding affinity of this substrate analogue (BODIPY-FL-ATPγS).
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0006-2960 1520-4995
DOI:	10.1021/acs.biochem.8b00485