Swiveling-domain mechanism for enzymatic phosphotransfer between remote reaction sites.

Herzberg, Osnat; Chen, Celia C. H.; Kapadia, Geeta; McGuire, Marielena; Carroll, Lawrence J.; Noh, Seong J.; Dunaway‐Mariano, Debra

doi:10.1073/pnas.93.7.2652

Cited by 144 publications

(219 citation statements)

References 33 publications

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“…In the HPr-binding domain that forms a four-helix bundle, one of the helices (residues 66-96) has a kink at residue 82, which as the NMR structure of EIN͞HPr complex shows (29), provides a pocket for HPr binding. The phosphorylated His-189 resides on the N terminus of one of the helices of the His domain, the domain that exhibits an ␣͞␤ fold unique to EI and PPDK (32). The polypeptide chain crosses over twice from the His domain to the HPr-binding domain such that the HPr-binding domain is inserted between two ␤-strands of the His domain.…”

Section: Results and Discussion Eimentioning

confidence: 99%

“…An ␣-helical domain binds HPr (the HPr-binding domain), and an ␣͞␤ domain bears the phosphorylation active center (the His domain). The His domain is functionally and structurally similar to the phosphohistidine swiveling domain of pyruvate phosphate dikinase (PPDK) (32).…”

mentioning

confidence: 99%

“…The structural and functional PPDK counterpart of EIC binds PEP and mediates dimerization as well. The EI Cys-502 counterparts in PPDK (and pyruvate kinase) were proposed to play a role in protonation of the PEP pyruvyl moiety after cleavage of the phosphoryl group (32).…”

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confidence: 99%

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Structure of phosphorylated enzyme I, the phosphoenolpyruvate:sugar phosphotransferase system sugar translocation signal protein

Teplyakov

Lim

Zhu

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

179

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sugar transport ͉ phosphorylation ͉ x-ray crystallography T he phosphoenolpyruvate (PEP):sugar phosphotransferase system (PTS) (1) catalyzes the synchronized uptake and phosphorylation of a number of carbohydrates in eubacteria (group translocation) (2, 3). With some variations, the PTS comprises three proteins. In the cytoplasm, PEP phosphorylates enzyme I (EI), which then transfers the phosphoryl group to the histidine phosphocarrier protein, HPr. From HPr, the phosphoryl group is transferred to various sugar-specific membrane associated transporters [enzyme II (EII)], each comprising two cytoplasmic domains, EIIA and EIIB, and an integral membrane domain EIIC. Within EII, EIIA accepts the phosphoryl group from HPr and donates it to EIIB, whereupon EIIC mediates sugar translocation. In addition to controlling sugar translocation, the phosphorylation state of PTS proteins is associated with regulation of metabolic pathways and signaling in bacterial cells (4-8).The Ϸ64-kDa EI is a homodimer, which is more tightly associated at the phosphorylated state than the unphosphorylated state (9-14). The phosphorylation by PEP requires Mg 2ϩ and targets the N atom of His-189 (numbering scheme of EI from Escherichia coli) (15). The dimer association rate constant is two to three orders of magnitude slower than typical rates measured for other dimeric proteins, suggesting that oligomerization is accompanied by major conformational rearrangements (13,16,17). The monomer-dimer equilibrium has been studied in vitro by various methods (18-21), and it has been proposed that the transition plays a regulatory role in the PEP:sugar phosphotransferase system. Yet, transient kinetic studies indicated that the EI dimer phosphorylates HPr without dissociating into monomers (17).Proteolytic cleavage of EI produces two domains (22, 23). The EI N-terminal domain (EIN, residues 1-230) contains the residue that transfers the phosphoryl group, 24) and the HPr-binding domain, whereas the EI C-terminal domain (EIC, residues 261-575) binds PEP in the presence of Mg 2ϩ (the PEP-binding domain) (22,25) and mediates dimerization (26,27). Site-directed mutagenesis showed that Cys-502, located on EIC, is essential for phosphorylation of His-189 by PEP (28). The structure of EIN from E. coli has been determined by x-ray crystallography (29) and NMR spectroscopy (30).

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Section: Results and Discussion Eimentioning

confidence: 99%

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confidence: 99%

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Structure of phosphorylated enzyme I, the phosphoenolpyruvate:sugar phosphotransferase system sugar translocation signal protein

Teplyakov

Lim

Zhu

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

179

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“…3A). The x-ray structure of maize PPDK reveals a three-domain structure consisting of a consecutive N-terminal ATP-grasp region, a central domain containing His-455 (His-529 in the full sequence of maize PPDK), and a C-terminal domain (Herzberg et al, 1996;McGuire et al, 1998). The identified phosphorylation sites at (1) Thr-309 and (2) Ser-506, Thr-527, and Ser-528 were located in the N-terminal ATP-grasp domain or the central His-529 PEP-utilizer domain, respectively (Fig.…”

Section: Identification Of Multiple Phosphorylation Sites On C 4 Ppdkmentioning

confidence: 99%

Posttranslational Modification of Maize Chloroplast Pyruvate Orthophosphate Dikinase Reveals the Precise Regulatory Mechanism of Its Enzymatic Activity

Chen

Wang

et al. 2014

Plant Physiol.

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In C 4 plants, pyruvate orthophosphate dikinase (PPDK) activity is tightly dark/light regulated by reversible phosphorylation of an active-site threonine (Thr) residue; this process is catalyzed by PPDK regulatory protein (PDRP). Phosphorylation and dephosphorylation of PPDK lead to its inactivation and activation, respectively. Here, we show that light intensity rather than the light/dark transition regulates PPDK activity by modulating the reversible phosphorylation at Thr-527 (previously termed Thr-456) of PPDK in maize (Zea mays). The amount of PPDK (unphosphorylated) involved in C 4 photosynthesis is indeed strictly controlled by light intensity, despite the high levels of PPDK protein that accumulate in mesophyll chloroplasts. In addition, we identified a transit peptide cleavage site, uncovered partial amino-terminal acetylation, and detected phosphorylation at four serine (Ser)/Thr residues, two of which were previously unknown in maize. In vitro experiments indicated that Thr-527 and Ser-528, but not Thr-309 and Ser-506, are targets of PDRP. Modeling suggests that the two hydrogen bonds between the highly conserved residues Ser-528 and glycine-525 are required for PDRP-mediated phosphorylation of the active-site Thr-527 of PPDK. Taken together, our results suggest that the regulation of maize plastid PPDK isoform (C 4 PPDK) activity is much more complex than previously reported. These diverse regulatory pathways may work alone or in combination to fine-tune C 4 PPDK activity in response to changes in lighting.

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“…The lysine which forms the Schiff's base with the PLP is predicted to be Lys-485 in PigH, and the aspartate which is hydrogen bonded to N-1 of PLP is predicted to be Asp-451. Herzberg et al, 1996). In PigC these two domains are separated by a domain that has no significant sequence identity to any protein of known function.…”

Section: Biosynthesis Of Mapmentioning

confidence: 99%

The Serratia gene cluster encoding biosynthesis of the red antibiotic, prodigiosin, shows species- and strain-dependent genome context variation

et al. 2004

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The prodigiosin biosynthesis gene cluster (pig cluster) from two strains of Serratia (S. marcescens ATCC 274 and Serratia sp. ATCC 39006) has been cloned, sequenced and expressed in heterologous hosts. Sequence analysis of the respective pig clusters revealed 14 ORFs in S. marcescens ATCC 274 and 15 ORFs in Serratia sp. ATCC 39006. In each Serratia species, predicted gene products showed similarity to polyketide synthases (PKSs), non-ribosomal peptide synthases (NRPSs) and the Red proteins of Streptomyces coelicolor A3(2). Comparisons between the two Serratia pig clusters and the red cluster from Str. coelicolor A3(2) revealed some important differences. A modified scheme for the biosynthesis of prodigiosin, based on the pathway recently suggested for the synthesis of undecylprodigiosin, is proposed. The distribution of the pig cluster within several Serratia sp. isolates is demonstrated and the presence of cryptic clusters in some strains shown. The pig cluster of Serratia marcescens ATCC 274 is flanked by cueR and copA homologues and this configuration is demonstrated in several S. marcescens strains, whilst these genes are contiguous in strains lacking the pig cluster.

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Swiveling-domain mechanism for enzymatic phosphotransfer between remote reaction sites.

Abstract: Fig. 1 (2, 3). Enzyme phosphorylation occurs on the N8 atom of a histidine residue (4-6), which has been identified as His-455 in the amino acid sequence of PPDK from Clostridium symbiosum, the subject of this study (6

Cited by 144 publications

References 33 publications

Structure of phosphorylated enzyme I, the phosphoenolpyruvate:sugar phosphotransferase system sugar translocation signal protein

Structure of phosphorylated enzyme I, the phosphoenolpyruvate:sugar phosphotransferase system sugar translocation signal protein

Posttranslational Modification of Maize Chloroplast Pyruvate Orthophosphate Dikinase Reveals the Precise Regulatory Mechanism of Its Enzymatic Activity

The Serratia gene cluster encoding biosynthesis of the red antibiotic, prodigiosin, shows species- and strain-dependent genome context variation

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