Selective killing of Klebsiella pneumoniae by 5-trifluoromethylthioribose. Chemotherapeutic exploitation of the enzyme 5-methylthioribose kinase.

Gianotti, Andréia; Tower, Paula A.; Sheley, Jared; Conte, P A; Spiro, C; Ferro, Adolph J.; Fitchen, John H.; Riscoe, Michael K.

doi:10.1016/s0021-9258(19)40124-5

Cited by 31 publications

(7 citation statements)

References 33 publications

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“…MTR kinase activity has been detected in a variety of bacteria, protozoa and plants [1,3,[28][29][30][31][32]. The enzyme has been partially purified from Enterobacter aerogenes [29], K. pneumoniae [4] and seeds of Lupinus luteus [28]. Several investigators estimated the molecular mass of native MTR kinase to be in the range of 70 kDa, based on gel-filtration analysis [4,28].…”

Section: Discussionmentioning

confidence: 99%

“…The clinical isolate of K. pneumoniae as well as the culture conditions used to maintain the organism for these studies have been described previously [4].…”

Section: Bacterial Strains and Culture Conditionsmentioning

confidence: 99%

“…For enzyme purification purposes, MTR kinase activity was measured essentially as described by Gianotti et al [4]. Briefly, enzyme samples (1-10 µl) were incubated for 1 h at 37 mC in 100 µl of reaction mixture containing 100 mM glycine (pH 9.5), 5 mM ATP, 20 mM MgCl # , 5 mM DTT and 5 µM "%C-MTR (0.029 µCi).…”

Section: Enzyme Assays and Kineticsmentioning

confidence: 99%

“…Because of its prevalence in a number of bacterial and protozoan pathogens of humans, attention has turned to the enzyme as a target for the development of drugs to selectively exploit or subvert microbial methionine metabolism [1][2][3]. It has been demonstrated that MTR kinase-containing organisms are selectively killed by analogues of MTR [3][4][5][6]. Depending on their design, these analogues kill microbes by one or more of the following mechanisms : (1) direct inhibition of MTR kinase activity, thus preventing salvage of the energetically ' expensive ' amino acid methionine ; (2) conversion of the drug via MTR kinase and the salvage pathway into toxic derivatives of methionine or S-adenosylmethionine (SAM) ; and (3) conversion via MTR kinase to unstable intermediates which yield toxic products upon decomposition.…”

Section: Introductionmentioning

confidence: 99%

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Affinity purification of 5-methylthioribose kinase and 5-methylthioadenosine/S-adenosylhomocysteine nucleosidase from Klebsiella pneumoniae

Cornell

Winter²,

Tower

et al. 1996

Biochemical Journal

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Two enzymes in the methionine salvage pathway, 5-methylthioribose kinase (MTR kinase) and 5'-methylthioadenosine/ S-adenosylhomocysteine nucleosidase (MTA/SAH nucleosidase) were purified from Klebsiella pneumoniae. Chromatography using a novel 5'-(p-aminophenyl)thioadenosine/5-(p-aminophenyl)thioribose affinity matrix allowed the binding and selective elution of each of the enzymes in pure form. The molecular mass, substrate kinetics and N-terminal amino acid sequences were characterized for each of the enzymes. Purified MTR kinase exhibits an apparent molecular mass of 46-50 kDa by SDS/PAGE and S200HR chromatography, and has a Km for MTR of 12.2 microM. Homogeneous MTA/SAH nucleosidase displays a molecular mass of 26.5 kDa by SDS/PAGE, and a Km for MTA of 8.7 microM. Comparisons of the N-terminal sequences obtained for each of the enzymes with protein-sequence databases failed to reveal any significant sequence similarities to known proteins. However, the amino acid sequence obtained for the nucleosidase did share a high degree of sequence similarity with the putative translation product of an open reading frame in Escherichia coli, thus providing a tentative identification of this gene as encoding an MTA/SAH nucleosidase.

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Section: Discussionmentioning

confidence: 99%

“…The clinical isolate of K. pneumoniae as well as the culture conditions used to maintain the organism for these studies have been described previously [4].…”

Section: Bacterial Strains and Culture Conditionsmentioning

confidence: 99%

Section: Enzyme Assays and Kineticsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Affinity purification of 5-methylthioribose kinase and 5-methylthioadenosine/S-adenosylhomocysteine nucleosidase from Klebsiella pneumoniae

Cornell

Winter²,

Tower

et al. 1996

Biochemical Journal

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“…However, the way by which the sulfur moiety is recycled remains unknown. To elucidate this pathway, we used an analogue of MTR, trifluoromethylthioribose (3F-MTR), which has been assayed as a possible drug lead for killing pathogens [ 10 ]. After transposon mutagenesis (see Materials and Methods), eleven 3F-MTR resistant clones were obtained (located in genes ykrW, ykrY and ykrT ).…”

Section: Resultsmentioning

confidence: 99%

Untitled

et al. 2001

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Metabolic characteristics and importance of the universal methionine salvage pathway recycling methionine from 5′‐methylthioadenosine

2009

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SummaryThe methionine salvage pathway, also called the 5 0 -methylthioadenosine (MTA) cycle, recycles the sulfur of MTA, which is a by-product in the biosyntheses of polyamine and the plant hormone ethylene. MTA is first converted to 5 0 -methylthioribose-1-phosphate either by MTA phosphorylase or the combined action of MTA nucleosidase and 5 0 -methylthioribose kinase. Subsequently, five additional enzymatic steps, catalyzed by four or five proteins, will form 4-methylthio-2-oxobutyrate, the deaminated form of methionine. The final transamination is achieved by transaminases active in the amino acid biosynthesis. This pathway is present with some variations in all types of organisms and seems to be designed for a quick removal of MTA achieved by high affinities of the first enzymes. During evolution some enzymes have attained additional functions, like a proposed role in nuclear mRNA processing by the aci-reductone dioxygenase. For others the function seems to be lost due to conditions in specific ecological niches, such as, presence of sulfur and/or absence of oxygen resulting in that, for example, Escherichia coli is lacking a functional pathway. The pathway is regulated as response to sulfur availability and take part in the regulation of polyamine synthesis. Some of the enzymes in the pathway show separate specificities in different organisms and some others are unique for groups of bacteria and parasites. Thus, promising targets for antimicrobial agents have been identified. Other medical topics to which this pathway has connections are cancer, apoptosis, and inflammatory response.

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Selective killing of Klebsiella pneumoniae by 5-trifluoromethylthioribose. Chemotherapeutic exploitation of the enzyme 5-methylthioribose kinase.

Cited by 31 publications

References 33 publications

Affinity purification of 5-methylthioribose kinase and 5-methylthioadenosine/S-adenosylhomocysteine nucleosidase from Klebsiella pneumoniae

Affinity purification of 5-methylthioribose kinase and 5-methylthioadenosine/S-adenosylhomocysteine nucleosidase from Klebsiella pneumoniae

Untitled

Metabolic characteristics and importance of the universal methionine salvage pathway recycling methionine from 5′‐methylthioadenosine

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