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

Olivia M. Chase, Adeline Espinasse, Kaelyn E. Wilke, Erin E. Carlson

Research output: Contribution to journalArticlepeer-review

3 Scopus citations

Abstract

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).

Original languageEnglish (US)
Pages (from-to)4368-4373
Number of pages6
JournalBiochemistry
Volume57
Issue number29
DOIs
StatePublished - Jul 24 2018

Bibliographical note

Funding Information:
*E-mail: carlsone@umn.edu. ORCID Erin E. Carlson: 0000-0001-8287-8893 Funding This work was supported by the University of Minnesota, the University of Minnesota National Institutes of Health (NIH; T32GM008347) Biotechnology Training Grant (A.E.), NIH Grant DP2OD008592, and an Indiana University Quantitative and Chemical Biology Training Fellowship (K.E.W.). Notes The authors declare no competing financial interest.

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