Properties of polyphosphate kinase prepared from Mycobacterium smegmatis

Suzuki, Hiroyasu; Kaneko, Tadaaki; Ikeda, Yōnosuke

doi:10.1016/0005-2744(72)90333-6

Cited by 15 publications

(5 citation statements)

References 14 publications

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“…Our results suggest that the over-expression of Rv1026 may lower polyphosphate levels in M. smegmatis cells in an indirect manner. It would putatively reduce the polyphosphate-synthesizing activities of the M. smegmatis PPK1 protein, by lowering its substrate (ATP) levels and increasing it product (ADP)-mediated inhibition; amongst other possible pleiotropic effects [8], [30], [39], [50]. However, we cannot exclude the possibility that Rv1026 functions as an exopolyphosphatase when expressed within M. tuberculosis or other mycobacterial cells.…”

Section: Discussionmentioning

confidence: 97%

The Two PPX-GppA Homologues from Mycobacterium tuberculosis Have Distinct Biochemical Activities

et al. 2012

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Inorganic polyphosphate (poly-P), guanosine pentaphosphate (pppGpp) and guanosine tetraphosphate (ppGpp) are ubiquitous in bacteria. These molecules play a variety of important physiological roles associated with stress resistance, persistence, and virulence. In the bacterial pathogen Mycobacterium tuberculosis, the identities of the proteins responsible for the metabolism of polyphosphate and (p)ppGpp remain to be fully established. M. tuberculosis encodes two PPX-GppA homologues, Rv0496 (MTB-PPX1) and Rv1026, which share significant sequence similarity with bacterial exopolyphosphatase (PPX) and guanosine pentaphosphate 5′-phosphohydrolase (GPP) proteins. Here we delineate the respective biochemical activities of the Rv0496 and Rv1026 proteins and benchmark these against the activities of the PPX and GPP proteins from Escherichia coli. We demonstrate that Rv0496 functions as an exopolyphosphatase, showing a distinct preference for relatively short-chain poly-P substrates. In contrast, Rv1026 has no detectable exopolyphosphatase activities. Analogous to the E. coli PPX and GPP enzymes, the exopolyphosphatase activities of Rv0496 are inhibited by pppGpp and, to a lesser extent, by ppGpp alarmones, which are produced during the bacterial stringent response. However, neither Rv0496 nor Rv1026 have the ability to hydrolyze pppGpp to ppGpp; a reaction catalyzed by E. coli PPX and GPP. Both the Rv0496 and Rv1026 proteins have modest ATPase and to a lesser extent ADPase activities. pppGpp alarmones inhibit the ATPase activities of Rv1026 and, to a lesser extent, the ATPase activities of Rv0496. We conclude that PPX-GppA family proteins may not possess all the catalytic activities implied by their name and may play distinct biochemical roles involved in polyphosphate and (p)ppGpp metabolic pathways.

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Section: Discussionmentioning

confidence: 97%

The Two PPX-GppA Homologues from Mycobacterium tuberculosis Have Distinct Biochemical Activities

et al. 2012

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“…However, in view of the delicate stability of the enzyme which resulted in low recovery, it cannot entirely be excluded that the 55-kDa protein represents a stabilized, active subunit of the holoenzyme. (8,9,19,20 About 50% of the total amount of P of the P5 was in the form of P2, P3, and P4.…”

Section: Discussionmentioning

confidence: 99%

Properties of polyphosphate: AMP phosphotransferase of Acinetobacter strain 210A

Bonting¹,

Kortstee²,

Zehnder³

1991

J Bacteriol

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Polyphosphate:AMP phosphotransferase, an enzyme which catalyzes the phosphorylation of AMP to ADP at the expense of polyphosphate, was purified more than 1,500-fold from Acinetobacter strain 210A by streptomycin sulfate precipitation and by Mono-Q, Phenyl Superose, and Superose column chromatography. Streptomycin sulfate precipitation appeared to be an effective step in the purification procedure. During the following chromatographic steps, there was a 29-fold increase in specific activity but the yield was low (0.3%).Kinetic studies showed apparent Km values of 0.26 mM for AMP and 0.8 PM for polyphosphate with an average chain length of 35 phosphate groups. The highest activities were found with polyphosphate molecules of 18 to 44 phosphate residues. The polyphosphate chain was degraded completely to ADP. The mechanism of degradation is processive. No activity was obtained with ortho-, pyro-, tri-, and tetraphosphate. The enzyme was inhibited by pyro-, tri-, and tetraphosphate. The inhibition by tri-and tetraphosphate was mixed with polyphosphate as a substrate. The inhibition constants for the dissociation of the enzyme-inhibitor complex and for the enzyme-inhibitor-substrate complex were 0.9 and 6.5 mM, respectively, for triphosphate and 0.7 and 1.5 mM, respectively, for tetraphosphate.

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“…Several biochemical reactions are known to be involved in microbial synthesis and degradation of polyphosphates. The enzymes catalyzing their biosynthesis are Polyp i kinase (ATP:polyPi phosphotransferase, EC 2.7.4.1) [7] and PolyPi:-3-phosphoglycerate kinase (1,3-diphosphoglycerate: polyPt phosphotransferase, EC 2.7.4.17) [3]. They are involved in the transfer of the energy-rich phosphoryl group from either ATP or 1,3-P2glycerate to PolyPl.…”

Section: Polyphosphate Metabolismmentioning

confidence: 99%

Studies on phosphate metabolism in obligate methanotrophs

Trotsenko

1990

FEMS Microbiology Letters

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SUMMARYStudies on three representative species of methane-ufflizing bacteria revealed high intracellular levels of inorganic pyrophosphate (5 mM) in contrast to low levels of ATP (0.5 mM) as well as the differences in the sets of enzymes utilizing pyro-and polypbosphates as phosphate and metabolic energy donors. INTRODUCTIONOver the last two decades there has been considerable progress in our understanding of the biochemistry of carbon and nitrogen metabolic pathways in aerobic C t utilizers. However, so far, surprisingly little is known about their phosphate metabolism, although the importance of phosphates in cellular global metabolic networks is well recognized. In an attempt to close this gap we have recently started investigations of phosphate metabolism in obligate methane utilizers. These bacteria are very promising for such studies since there is a good correlation between the different types of their abundant intracellular membranes, consisting mainly of phospbolipids, and carbon Correspondence to: Y.A. Trotsenko, Institute of Biochemistry ",.~' Pl~ysiolo~," ~f ~.~,,6organisms, U.S.S.R. Academy of Sciences, Pushchlno, 142292, U.S.S.R.and nitrogen assimilatory pathways [1,2]. It appears interesting and useful to investigate the integration of phosphate metabolism in general metabolic networks and their regulatory patterns in various types of methanotrophs.Our studies on phosphate metabolism in obligate methanotrophs have concentrated on (1) the estimation of intracellular pools of inorganic orthophosphate (Pi), pyropbosphate (PPi) and polyphosphates (Polyp i), (2) the determination of the activity levels of phosphate-metabolising enzymes, as well as (3) the regulatory aspects of phosphate metabolism. Three representative species of methanotrophs were used throughout the investigations: Methylomonas methanica, a Type I organism, having the ribulosemonophosphate (RUMP) cycle, an incomplete tricarboxyfic acid (TCA) cycle and glutamate dehydrogenase, Methylosinus trichosporium (Type II; serine cycle; complete TCA cycle; glutamate cycle) and the 'mosaic organism' Methylococcus capsulatus (Type X; RuMP plus ribulosebisphosphate (RuBP) cycles; alanine dehydrogenase plus glutamate cycle).As mentioned above, our attention has been focused on the metabolism of inorganic pyro-and polyphosphates which are known to be linear polymers of orthophosphate in anhydrous linkage. Their length, expressed by the number of phosphate residues in the chain, may range from two residues, as in PPi, to more than 1000 residues, as in PolyPis. These compounds with the high-energy pbosphoanhydride bonds are widely distributed in 0168-6445/90/$03.50

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Properties of polyphosphate kinase prepared from Mycobacterium smegmatis

Cited by 15 publications

References 14 publications

The Two PPX-GppA Homologues from Mycobacterium tuberculosis Have Distinct Biochemical Activities

The Two PPX-GppA Homologues from Mycobacterium tuberculosis Have Distinct Biochemical Activities

Properties of polyphosphate: AMP phosphotransferase of Acinetobacter strain 210A

Studies on phosphate metabolism in obligate methanotrophs

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