5A8G

Crystal structure of the wild-type Staphylococcus aureus N- acetylneurminic acid lyase in complex with fluoropyruvate

Summary for 5A8G

• Entry DOI: 10.2210/pdb5a8g/pdb
• Descriptor: N-ACETYLNEURAMINATE LYASE (2 entities in total)
• Functional Keywords: lyase
• Biological source: STAPHYLOCOCCUS AUREUS SUBSP. AUREUS NCTC 8325
• Cellular location: Cytoplasm : Q2G160
• Total number of polymer chains: 2
• Total formula weight: 68242.97

Authors

Stockwell, J.,Daniels, A.D.,Windle, C.L.,Harman, T.,Woodhall, T.,Trinh, C.H.,Lebel, T.,Pearson, A.R.,Mulholland, K.,Berry, A.,Nelson, A. (deposition date: 2015-07-15, release date: 2015-11-18, Last modification date: 2024-11-06)

Primary citation

Stockwell, J.,Daniels, A.D.,Windle, C.L.,Harman, T.A.,Woodhall, T.,Lebl, T.,Trinh, C.H.,Mulholland, K.,Pearson, A.R.,Berry, A.,Nelson, A.
Evaluation of Fluoropyruvate as Nucleophile in Reactions Catalysed by N-Acetyl Neuraminic Acid Lyase Variants: Scope, Limitations and Stereoselectivity.
Org.Biomol.Chem., 14:105-, 2016

PubMed Abstract: The catalysis of reactions involving fluoropyruvate as donor by N-acetyl neuraminic acid lyase (NAL) variants was investigated. Under kinetic control, the wild-type enzyme catalysed the reaction between fluoropyruvate and N-acetyl mannosamine to give a 90 : 10 ratio of the (3R,4R)- and (3S,4R)-configured products; after extended reaction times, equilibration occurred to give a 30 : 70 mixture of these products. The efficiency and stereoselectivity of reactions of a range of substrates catalysed by the E192N, E192N/T167V/S208V and E192N/T167G NAL variants were also studied. Using fluoropyruvate and (2R,3S)- or (2S,3R)-2,3-dihydroxy-4-oxo-N,N-dipropylbutanamide as substrates, it was possible to obtain three of the four possible diastereomeric products; for each product, the ratio of anomeric and pyranose/furanose forms was determined. The crystal structure of S. aureus NAL in complex with fluoropyruvate was determined, assisting rationalisation of the stereochemical outcome of C-C bond formation.

PubMed: 26537532
DOI: 10.1039/C5OB02037A

Experimental method

X-RAY DIFFRACTION (1.72 Å)