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Updated: Sep 25, 2026

Quantification of Bacterial Histidine Kinase Autophosphorylation Using a Nitrocellulose Binding Assay
Published on: January 11, 2017
ATP-citrate lyase as a substrate of protein histidine phosphatase in vertebrates
Susanne Klumpp1, Gunther Bechmann, Anette Mäurer
1Institut für Pharmazeutische and Medizinische Chemie, Westfälische Wilhelms-Universität, Hittorfstr. 58-62, D-48149, Münster, Germany. klumpp@uni-muenster.de
Abstract:
The first protein histidine phosphatase from vertebrates discovered recently was found in a variety of tissues, however, a physiological substrate protein was missing. Phosphorylation of liver extracts in the presence of EDTA, followed by SDS-PAGE and autoradiography showed labeling of three proteins. Acid- and alkaline-treatment revealed the existence of N-phosphates. Addition of histidine phosphatase exclusively resulted in dephosphorylation of a 110kDa protein (denaturing conditions). Gelfiltration revealed its native molecular mass of approximately 450kDa. That protein was purified and identified as ATP-citrate lyase. The results are in favor of histidine phosphatase playing an important yet unidentified role in metabolic processes.
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