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Specificity of the udp n acetylmuramyl l alanyl d glutamate meso 2 6 diaminiopimelate synthetase from escherichia coli


, : Specificity of the udp n acetylmuramyl l alanyl d glutamate meso 2 6 diaminiopimelate synthetase from escherichia coli. European Journal of Biochemistry 153(1): 81-88

To investigate the specificity of the uridine-diphosphate-N-acetylmuramyl-L-alanyl-D-glutamate:meso-2,6-diaminopimelate synthetase, various compounds mimicking more or less different parts of the UDP-MurNAc-L-Ala-D-Glu substrate were prepared. Their size ranged from that of uridine or L-Ala-D-Glu to that of the whole nucleotide substrate. Chemical synthesis led to N.alpha.-acyl-dipeptides, in which the acyl group mimicked the MurNAc moiety, and to glycopeptides MurNac(.alpha. or .beta.-Me)-L-Ala-D-Glu, in which the anomeric function is blocked. Partial degradation or chemical modification of the substrate UDP-MurNAc-L-Ala-D-Glu afforded: MurOHNAc-L-Ala-D-Glu, P1-MurNAc-L-Ala-D-Glu, and DDP-MurNAc-L-Ala-D-Glu (DDP = dihydrouridine-diphosphate). All these compounds were tested as substrates or (and) inhibitors of the reaction catalyzed by the A2pm-adding enzyme, which, after partial purification, was obtained in two active forms. Among the compounds tested as substrates, only DDP-MurNAc-L-Ala-D-Glu was a good one. The Km for this compound was 97 .mu.M versus 55 .mu.M for the natural substrate. Among the various compounds tested as inhibitors, only P1-MurNAc-L-Ala-D-Glu and MurNAc(.alpha. or .beta.-Me)-L-Ala-D-Glu had a significant inhibitory effect at 1 mM. Apparently, no particular portion of the molecule is predominantly responsible for its recognition by the enzyme. In other words, multiple sites located over the whole molecule are required for a proper recognition and determine the high specificity of this activity. Therefore, to obtain efficient competitive inhibitors it is necessary to synthesize molecules very similar in size and structure to the natural substrate.

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Related references

Abo-Ghalia, M.; Michaud, C.; Blanot, D.; van Heijenoort, J., 1985: Specificity of the uridine-diphosphate-N-acetylmuramyl-L-alanyl-D-glutamate: meso-2,6-diaminopimelate synthetase from Escherichia coli. To investigate the specificity of the uridine-diphosphate-N-acetylmuramyl-L-alanyl-D-glutamate: meso-2,6-diaminopimelate synthetase, various compounds mimicking more or less different parts of the UDP-MurNAc-L-Ala-D-Glu substrate were prepared. Th...

Mizuno, Y.; Ito, E., 1968: Purification and properties of uridine diphosphate N-acetylmuramyl-L-alanyl-D-glutamate:meso-2,6-diaminopimelate ligase. Journal of Biological Chemistry 243(10): 2665-2672

Anonymous, 1968: Purification and properties of uridine diphosphate N-acetylmuramyl-L-alanyl-D-glutamate Meso-2,6-diaminopimelate ligase

Gordon, E.; Flouret, B.; Chantalat, L.; van Heijenoort, J.; Mengin-Lecreulx, D.; Dideberg, O., 2001: Crystal structure of UDP-N-acetylmuramoyl-L-alanyl-D-glutamate: meso-diaminopimelate ligase from Escherichia coli. UDP-N-acetylmuramoyl-l-alanyl-d-glutamate:meso-diaminopimelate ligase is a cytoplasmic enzyme that catalyzes the addition of meso-diaminopimelic acid to nucleotide precursor UDP-N-acetylmuramoyl-l-alanyl-d-glutamate in the biosynthesis of bacteria...

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Michaud, C.; Mengin-Lecreulx, D.; van Heijenoort, J.; Blanot, D., 1990: Over-production, purification and properties of the uridine-diphosphate-N-acetylmuramoyl-L-alanyl-D-glutamate: meso-2,6-diaminopimelate ligase from Escherichia coli. The UDP-N-acetylmuramoyl-L-alanyl-D-glutamate:meso-2,6-diaminopimelate ligase was over-produced and purified from two plasmid-harbouring strains of Escherichia coli. The first strain, E. coli JM83(pHE5), gave a 15-fold over-production relative to...

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