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Contribution of the active site aspartic acid to catalysis in the bacterial neuraminidase from Micromonospora viridifaciens.

Watson JN, Newstead S, Dookhun V, Taylor G, Bennet AJ
FEBS Lett 577 265-9 (2004)
Cited: 10 times
EuropePMC logo PMID: 15527797

Abstract

A recombinant D92G mutant sialidase from Micromonospora viridifaciens has been cloned, expressed and purified. Kinetic studies reveal that the replacement of the conserved aspartic acid with glycine results in a catalytically competent retaining sialidase that possesses significant activity against activated substrates. The contribution of this aspartate residue to the free energy of hydrolysis for natural substrates is greater than 19 kJ/mol. The three dimensional structure of the D92G mutant shows that the removal of aspartic acid 92 causes no significant re-arrangement of the active site, and that an ordered water molecule substitutes for the carboxylate group of D92.

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  1. Sequence and structural analysis of the Asp-box motif and Asp-box beta-propellers; a widespread propeller-type characteristic of the Vps10 domain family and several glycoside hydrolase families. Quistgaard EM, Thirup SS. BMC Struct Biol 9 46 (2009)
  2. It All Starts with a Sandwich: Identification of Sialidases with Trans-Glycosylation Activity. Nordvang RT, Nyffenegger C, Holck J, Jers C, Zeuner B, Sundekilde UK, Meyer AS, Mikkelsen JD. PLoS One 11 e0158434 (2016)


Reviews citing this publication (1)

  1. Features and applications of bacterial sialidases. Kim S, Oh DB, Kang HA, Kwon O. Appl Microbiol Biotechnol 91 1-15 (2011)

Articles citing this publication (7)

  1. Decreasing the sialidase activity of multifunctional Pasteurella multocida α2-3-sialyltransferase 1 (PmST1) by site-directed mutagenesis. Sugiarto G, Lau K, Li Y, Khedri Z, Yu H, Le DT, Chen X. Mol Biosyst 7 3021-3027 (2011)
  2. Computational modeling of laminin N-terminal domains using sparse distance constraints from disulfide bonds and chemical cross-linking. Kalkhof S, Haehn S, Paulsson M, Smyth N, Meiler J, Sinz A. Proteins 78 3409-3427 (2010)
  3. Natural sialoside analogues for the determination of enzymatic rate constants. Indurugalla D, Watson JN, Bennet AJ. Org Biomol Chem 4 4453-4459 (2006)
  4. Two nucleophilic mutants of the Micromonospora viridifaciens sialidase operate with retention of configuration by two different mechanisms. Watson JN, Newstead S, Narine AA, Taylor G, Bennet AJ. Chembiochem 6 1999-2004 (2005)
  5. Hydrophilic aromatic residue and in silico structure for carbohydrate binding module. Chou WY, Pai TW, Jiang TY, Chou WI, Tang CY, Chang MD. PLoS One 6 e24814 (2011)
  6. Use of conformationally restricted pyridinium alpha-D-N-acetylneuraminides to probe specificity in bacterial and viral sialidases. Watson JN, Knoll TL, Chen JH, Chou DT, Borgford TJ, Bennet AJ. Biochem Cell Biol 83 115-122 (2005)
  7. Molecular Dynamics Simulations Reveal the Conformational Transition of GH33 Sialidases. Cao X, Yang X, Xiao M, Jiang X. Int J Mol Sci 24 6830 (2023)


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