Dataset Information

Chemoproteomic identification of a phosphohistidine acceptor: Insights into posttranslational regulation of glycolysis

ABSTRACT: Histidine phosphorylation is an underexplored form of protein phosphorylation. Despite the widespread existence of phosphohistidine (pHis) sites, particularly in bacteria, their phosphorylation regulation and physiological functions remain poorly understood. In this study, we developed a chemoproteomic strategy employing a stable pHis analog to identify pHis-recognizing proteins in Escherichia coli. Our probe successfully labeled known pHis-recognizers and revealed many putative pHis acceptors, including phosphofructokinase-1 (PfkA), a key glycolytic enzyme. We demonstrated that PfkA undergoes histidine phosphorylation at His249 mediated by the phosphocarrier protein PtsH, thereby reducing enzyme activity. This phosphorylation was reversed by the pHis-specific phosphatase SixA, which restored the PfkA activity. Our findings reveal a novel posttranslational regulatory mechanism affecting glycolysis, implicating a broader role of histidine phosphorylation in bacterial metabolic control.

INSTRUMENT(S):

ORGANISM(S): Escherichia Coli

TISSUE(S): Cell Culture

SUBMITTER: Seungmin Ahn

LAB HEAD: Jung-Min Kee

PROVIDER: PXD053006 | Pride | 2025-05-07

REPOSITORIES: Pride

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Dataset's files

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			Action	DRS
	Me-BP_1_20230221_QE_630_23FE22_AhnSM.raw	Raw
	Me-BP_2_20230221_QE_630_23FE22_AhnSM.raw	Raw
	Me-BP_3_20230221_QE_630_23FE22_AhnSM.raw	Raw
	PhosphoSTYHSites_20221228_QE_617_22DE28_AhnSM.txt	Txt
	Supplementary3_20221109_QE_609_22NO09_AhnSM.raw	Raw

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Chemoproteomic identification of phosphohistidine acceptors: posttranslational activity regulation of a key glycolytic enzyme.

Choi Solbee S Ahn Seungmin S Cho Kyung Hyun KH Lee Sung Kuk SK Kee Jung-Min JM

Chemical science 20250331 18

Histidine phosphorylation, an unconventional and understudied posttranslational modification, often involves phosphohistidine (pHis) "acceptor" proteins, which bind to pHis residues and undergo phosphotransfer from pHis. While the roles of pHis acceptors are well-documented in bacterial cell signalling and metabolism, the presence and functions of additional pHis acceptors remain largely unknown. In this study, we introduce a chemoproteomic strategy leveraging a stable analogue of 3-pHis to iden ...[more]

PMID: 40201162

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