Mechanism of Dephosphorylation of Glucosyl-3-phosphoglycerate by a Histidine Phosphatase

Qin, Zheng; Jiang, Dunquan; Zhang, Wei; Zhang, Qingqing; Zhao, Qi; Jin, Jin; Li, Xin; Yang, Haitao; Bartlam, Mark; Shaw, Neil; Zhou, Weihong; Rao, Zihe

doi:10.1074/jbc.m114.569913

Cited by 10 publications

(9 citation statements)

References 48 publications

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“…The phosphatase activity of Sts-1, like that of other PGM family histidine phosphatases, is proposed to proceed through a twostep mechanism that involves nucleophilic attack by one of the conserved histidine residues. 6,8,[18][19][20]30 The active site glutamate is the likely proton donor during this attack. The second step of the reaction is a hydrolysis of the phosphor−histidine intermediate.…”

Section: ■ Results and Discussionmentioning

confidence: 99%

“…Functional studies, both in vitro and in vivo , showed that Sts-1 has a much higher phosphatase activity than that of Sts-2, both on non-native substrates and the putative protein substrate, Zap-70 6, 11, 14, 17 . The mechanism of catalysis is believed to resemble that of other histidine phosphatases, proceeding through a covalent phosphor-histidine intermediate 8, 18–20 .…”

Section: Introductionmentioning

confidence: 99%

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Structural and Functional Characterization of the Histidine Phosphatase Domains of Human Sts-1 and Sts-2

et al. 2017

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The suppressor of T-cell signaling (Sts) proteins, Sts-1 and Sts-2, are homologous phosphatases that negatively regulate signaling pathways downstream of the T-cell receptor. Functional inactivation of Sts-1 and Sts-2 in a murine model leads to resistance to systemic infection by the opportunistic pathogen, C. albicans. This suggests that modulation of the host immune response by inhibiting Sts function may be a viable strategy to treat these deadly fungal pathogen infections. To better understand the molecular determinants of function and structure, we characterized the structure and steady-state kinetics of the histidine phosphatase domains of human Sts-1 (Sts-1HP) and Sts-2 (Sts-2HP). We solved the X-ray crystal structures of Sts-1HP, unliganded and in complex with sulfate to 2.5 Å and 1.9 Å, respectively, and the structure of Sts-2HP with sulfate to 2.4 Å. The steady-state kinetic analysis shows, as expected, that Sts-1HP has a significantly higher phosphatase activity than that of Sts-2HP, and that the human and mouse proteins behave similarly. In addition, comparison of the phosphatase activity of full-length Sts-1 protein to Sts-1HP reveals similar kinetics, indicating that Sts-1HP is a functional surrogate for the native protein. We also tested known phosphatase inhibitors and identified that the SHP-1 inhibitor, PHPS1, is a potent inhibitor of Sts-1 (Ki of 1.05 ± 0.15 μM). Finally, we demonstrated that human Sts-1 has robust phosphatase activity against the substrate, Zap-70, in a cell-based assay. Collectively, these data suggest that the human Sts proteins are druggable targets and provides a structural basis for future drug development efforts.

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Section: ■ Results and Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Structural and Functional Characterization of the Histidine Phosphatase Domains of Human Sts-1 and Sts-2

et al. 2017

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“…The second step in the MGLP pathway was found to be catalyzed by an atypical glucosyl-3-phosphoglycerate phosphatase (GpgP, Rv2419c in M. tuberculosis H37Rv) of the histidine phosphatase superfamily that was not a sequence homolog of the isofunctional GpgPs of the HAD superfamily (EC 3.1.3.85), initially identified in other microorganisms able to synthesize GG132140. After their identification, these two MGLP enzymes have been crystallized and their three-dimensional structures determined for their potential as TB targets4142. MGLP seems to be ubiquitous among mycobacteria and GpgS and GpgP genes are present in all their available genomes (Table 1).…”

Section: Discussionmentioning

confidence: 99%

Mycobacterium hassiacum recovers from nitrogen starvation with up-regulation of a novel glucosylglycerate hydrolase and depletion of the accumulated glucosylglycerate

Alarico

Costa

Sousa

et al. 2014

Sci Rep

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Some microorganisms accumulate glucosylglycerate (GG) during growth under nitrogen deprivation. However, the molecular mechanisms underlying the role of GG and the regulation of its levels in the nitrogen stress response are elusive. Since GG is required for biosynthesis of mycobacterial methylglucose lipopolysaccharides (MGLP) we examined the molecular mechanisms linking replenishment of assimilable nitrogen to nitrogen-starved M. hassiacum with depletion of GG accumulated during nitrogen deficiency. To probe the involvement of a newly identified glycoside hydrolase in GG depletion, we produced the mycobacterial enzyme recombinantly and confirmed the specific hydrolysis of GG (GG hydrolase, GgH) in vitro. We have also observed a pronounced up-regulation of GgH mRNA in response to the nitrogen shock, which positively correlates with GG depletion in vivo and growth stimulation, implicating GgH in the recovery process. Since GgH orthologs seem to be absent from most slowly-growing mycobacteria including M. tuberculosis, the disclosure of the GgH function allows reconfiguration of the MGLP pathway in rapidly-growing species and accommodation of this possible regulatory step. This new link between GG metabolism, MGLP biosynthesis and recovery from nitrogen stress furthers our knowledge on the mycobacterial strategies to endure a frequent stress faced in some environments and during long-term infection.

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“…gordonae ) comprising the glucosyl-3-phosphoglycerate phosphatase domain is shown in the supplementary data ( S3 Appendix ). Some of these PE proteins, for instance, WP_003403850.1, WP_015357328.1 and CPR12073.1 recognized the template PDB_ID: 4PZ9:B corresponding to the mycobacterial glucosyl-3-phosphoglycerate phosphatase Rv2419c [ 55 ]. This enzyme consists of a single domain made up of a central β-sheet flanked by α-helices on either side and is known to catalyze the second step in the biosynthesis of methylglucose lipopolysaccharides (MGLPs) pathway.…”

Section: Resultsmentioning

confidence: 99%

Prediction of Certain Well-Characterized Domains of Known Functions within the PE and PPE Proteins of Mycobacteria

et al. 2016

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The PE and PPE protein family are unique to mycobacteria. Though the complete genome sequences for over 500 M. tuberculosis strains and mycobacterial species are available, few PE and PPE proteins have been structurally and functionally characterized. We have therefore used bioinformatics tools to characterize the structure and function of these proteins. We selected representative members of the PE and PPE protein family by phylogeny analysis and using structure-based sequence annotation identified ten well-characterized protein domains of known function. Some of these domains were observed to be common to all mycobacterial species and some were species specific.

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Mechanism of Dephosphorylation of Glucosyl-3-phosphoglycerate by a Histidine Phosphatase

Cited by 10 publications

References 48 publications

Structural and Functional Characterization of the Histidine Phosphatase Domains of Human Sts-1 and Sts-2

Structural and Functional Characterization of the Histidine Phosphatase Domains of Human Sts-1 and Sts-2

Mycobacterium hassiacum recovers from nitrogen starvation with up-regulation of a novel glucosylglycerate hydrolase and depletion of the accumulated glucosylglycerate

Prediction of Certain Well-Characterized Domains of Known Functions within the PE and PPE Proteins of Mycobacteria

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