2-Keto-3-deoxy-6-phosphogluconic aldolase

Mavridis, I.M., et al. Structure of 2-keto-3-deoxy-6-phosphogluconate aldolase at 2.8 A resolution. 

Fong, S., Machajewski, T.D., Mak, C.C. and Wong, C.-H. (2000) Directed evolution of D-2-keto-3-deoxy-6-phosphogluconate aldolase to new variants for the efficient synthesis of d- and L-sugars. Chemistry Biology, 7, 873-883. 

The 2-keto-3-deoxy-6-phosphogluconate aldolase (KDPG aldolase EC 4.1.2.14) catalyzes the cleavage of the dehydration product of 6-phosphogluconate, (KDPG), into glyceraldehyde-3-phosphate and pyruvate in the Entner-Doudoroff pathway (Scheme 7, R = P03H2). This... 

H. P. Meloche, J. M. Ingram, and W. A. Wood, 2-Keto-3-deoxy-6-phosphogluconic aldolase (crystalline). Methods Enzymol. 9 520 (1966). 

L. Pocivavsek, C. A. Fierke, E. J. Toone, and J. H. Naismith, Directed evolution of a new catalytic site in 2-keto-3-deoxy-6-phosphogluconate aldolase from Escherichia coli, Structure 2001, 9, 1-10. 

M. C. Shelton, I. C. Cotterill, S. T. A. Novak, R. M. Poonawala, S. Sudarshan, and E. J. Toone, 2-Keto-3-deoxy-6-phosphogluconate aldolases as catalysts for stereocon-trolled carbon-carbon bond formation, J. Am. Chem. Soc., 118 (1996) 2117-2125. 

Enzyme used in the assays was purified from Azotobacter vinelandii A modification of the assay was used to follow the approach to equilibrium of the 2-keto-3-deoxy-6-phosphogluconate aldolase reaction. 

As mentioned, the mildness of NaBHaCN (coupled with its effectiveness and stability in aqueous media) has attracted considerable interest for applications in biochemical areas. Examples include the trapping of suspected imine intermediates produced in enzyme (mitochondrial monoamine oxidase) inactivation by amines, the establishment by reduction of the positions of imine-forming amines in 2-keto-3-deoxy-6-phosphogluconate aldolase, and the transfer labeling of methionyl-tRNA synthetase and methionyl-tRNA transformalase by treatment with periodate-treated tRNA. In fact, most biochemical applications of NaBHaCN have utilized in situ imine formation-reduction (i.e. reductive amination) conditions and will be further discussed in Section 1.2.2.3.1. 

Lebioda, L., Hatada, M. H., Tulinsky, A., and Mavridis, I. Comparison of the folding of 2-keto-3-deoxy-6-phosphogluconate aldolase, triose phosphate iso-mercise and pyruvate kincise. Implications in molecular evolution. J. Molec. Biol. 162, 445-458 (1982). 

Keto-3-deoxy-6-phosphogluconate aldolase (KDPG aldolase) 165... 

Schiirmann M, Schiirmann M, Sprenger GA (2002) Fructose 6-phosphate aldolase and 1-deoxy-D-xylulose 5-phosphate synthase from Escherichia coli as tools in enzymatic synthesis of 1-deoxysugars. J Mol Catal B Enzym 19 247-252 Shelton CM, Toone EJ (1995) Differential dye-ligand chromatography as a general purification protocol for 2-keto-3-deoxy-6-phosphogluconate aldolases. Tetrahed Asymm 6 207-211 Silvestri MG, Desantis G, Mitchell M et al. (2003) Asymmetric aldol reactions using aldolases. Top Stereochem 23 267-342... 

The natural substrates for 2-keto-3-deoxy-6-phosphogluconate aldolase are D-glyceraldehyde-3-phosphate and pyruvate. It has now been shown that the enzyme will accept various other aldehydes as substrates, including D-glyceraldehyde itself, and D-lactaldehyde, from which the product is the dideoxyhexulosonate 25.29... 

Formation of a carbinolamine as the short-lived Schiff base precursor (C) could be trapped by Hash freezing of 2-keto-3-deoxy-6-phosphogluconate aldolase in the presence of its natural donor substrate pyruvate [46]. Stereospecificity of the reaction seems to be ensured mostly by hydrophobic contact of the pyruvate methyl group in the active site. 

There are 14 known occurrences of this motif but only six of these were present in the release of the Protein Data Bank used in this work enolase (lENL), 2-keto-3-deoxy-6-phosphogluconate aldolase (IKGA), pyruvate kinase (IPYK), triose phosphate isomerase (ITIM), D-xylose isomerase (IXIA) and taka-amylase (2TAA). The p-barrel from sub-unit 1 of ITIM was used to define the motif, and all distances and angles were employed in the search, together with the specification... 

Cotterill, I. C., Shelton, M. C., Machemer, D. E. W., Henderson, D. R, and Toone, E. J., Effect of phosphorylation on the reaction rate of unnatural electrophiles with 2-keto-3-deoxy-6-phosphogluconate aldolase.. Chem. Soc. Perkin Trans. 1998,1 (7), 1335-1342. 

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