The First Total Synthesis of Bacterial Cell Wall Precursor UDP−<i>N</i>-Acetylmuramyl-Pentapeptide (Park Nucleotide)

Hitchcock, Stephen A.; Eid, Clark N.; Aikins, James; Zia‐Ebrahimi, Mohammad; Blaszczak, Larry C.

doi:10.1021/ja973172d

Cited by 56 publications

(64 citation statements)

References 23 publications

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“…To date, several lipid II analogs have been synthesized and applied to the biochemical studies. [4] We have recently identified that neryl lipid II- N ε -dansyl analogue, 2 - N ε -dansylthiourea, could be recognized by mycobacterial transglycosylase (TGase) to form polymerized products with significant enhancement of visible-light absorption at 400 nm. Because the dansyl group itself does not have absorption in visible-light region, bathochromic shift of the polymerized- 2 - N ε -dansylthiourea is an unusual physicochemical observation.…”

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confidence: 99%

One‐Pot Protection‐Glycosylation Reactions for Synthesis of Lipid II Analogues

Mitachi

Mohan

Siricilla

et al. 2014

Chemistry A European J

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(2,6-Dichloro-4-methoxyphenyl)(2,4-dichlorophenyl) methyl trichloroacetimidate (3) and its polymer-supported reagent 4 can be successfully applied to a one-pot protection-glycosylation reaction to form the disaccharide derivative 7d for the synthesis of lipid II analogues. The temporary protecting group or linker at the C-6 position and N-Troc protecting group of 7d can be cleaved simoultaneousely via a reductive condition. Overall yield of syntheses of lipid II, 1 and neryl-lipid II-Nε-dansylthiourea are significantly improved via the described methods.

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confidence: 99%

One‐Pot Protection‐Glycosylation Reactions for Synthesis of Lipid II Analogues

Mitachi

Mohan

Siricilla

et al. 2014

Chemistry A European J

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“…Fractions containing UDPMurNAc-pentapeptide were identified by TLC on silica gel plates in 2-butyric acid/1 M NH 4 OH (5:3), utilizing UV absorption and ninhydrin for detection. These fractions were pooled and lyophilized to remove buffer, and the final product, UDP-MurNAc-pentapeptide, was analyzed by mass spectrometry as described (11,12). In most syntheses, the rate of conversion of UDP-MurNAc to UDP-MurNAc-pentapeptide was about 80%.…”

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confidence: 99%

Polymerization of Mycobacterial Arabinogalactan and Ligation to Peptidoglycan

Yagi¹,

Mahapatra

Mikušová³

et al. 2003

Journal of Biological Chemistry

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The cell wall of Mycobacterium spp. consists predominately of arabinogalactan chains linked at the reducing ends to peptidoglycan via a P-GlcNAc-(␣1-3)-Rha linkage unit (LU) and esterified to a variety of mycolic acids at the nonreducing ends. Several aspects of the biosynthesis of this complex have been defined, including the initial formation of the LU on a polyprenyl phosphate (Pol-P) molecule followed by the sequential addition of galactofuranosyl (Galf) units to generate Pol-P-P-LU-(Galf) 1,2,3, etc. and Pol-P-P-LU-galactan, catalyzed by a bifunctional galactosyltransferase (Rv3808c) capable of adding alternating 5-and 6-linked Galf units. By applying cell-free extracts of Mycobacterium smegmatis, containing cell wall and membrane fragments, and differential labeling with UDP-[ 14 C]Galp and recombinant UDP-Galp mutase as the source of [ C]Galf for galactan biosynthesis and 5-P-[14 C]ribosyl-P-P as a donor of [ 14 C]Araf for arabinan synthesis, we now demonstrate sequential synthesis of the simpler Pol-P-P-LU-(Galf) n glycolipid intermediates followed by the Pol-P-P-LU-arabinogalactan and, finally, ligation of the P-LU-arabinogalactan to peptidoglycan. This first time demonstration of in vitro ligation of newly synthesized P-LUarabinogalactan to newly synthesized peptidoglycan is a necessary forerunner to defining the genetics and enzymology of cell wall polymer-peptidoglycan ligation in Mycobacterium spp. and examining this step as a target for new antibacterial drugs.The cell envelope of Mycobacterium tuberculosis is composed of a conventional plasma membrane and a cell wall proper unique to some genera within the Actinomycetales order, consisting of a core of arabinogalactan (AG), 1 peptidoglycan (PG), and mycolic acids interspersed with a variety of free lipids, lipoglycans, and proteins (1); there is also evidence for polysaccharides on the outer face of the cell wall (2). The mycolic acids are attached to the nonreducing ends of the arabinogalactan, whereas the reducing ends are covalently attached to the crosslinked peptidoglycan via phosphoryl-N-acetylglucosaminosylrhamnosyl linkage units (P-GlcNAc-Rha). This massive structure, the mycolate-arabinogalactan-peptidoglycan-complex (MAPc), is the basis of many of the physiological and pathogenic features of M. tuberculosis and the site of susceptibility and resistance to many of the anti-tuberculosis drugs (3). Biosynthesis of this complex commences with attachment of the residues of the linkage unit, GlcNAc-1-P and Rha, donated by UDP-GlcNAc and dTDP-Rha, respectively, to a polyprenyl phosphate (Pol-P) carrier lipid (4). Formation of the linkage unit is followed by the sequential addition of galactofuranosyl (Galf) units donated by UDP-Galf, to provide simple Pol-P-Plinked AG intermediates (4). The bulk, if not all, of galactan biosynthesis is catalyzed by a membrane-associated bifunctional galactosyltransferase capable of adding the alternating 5-and 6-linked Galf units (5, 6).The demonstration that the direct donor of the arabinofuranosyl (Araf) unit...

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“…New assays cap- able of high-throughput screening of inhibitors targeting the transglycosylase are a subject of current interest. Chemical [41] and chemoenzymatic [42] syntheses of the Park nucleotide as well as the recent total syntheses of lipid II [43,44] and analogues with fluorescent labels facilitated the kinetic characterization of PBP1b and the development of a continuous fluorescence assay [45].…”

Section: Peptidoglycanmentioning

confidence: 99%