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. 2010 Nov;66(Pt 11):1257-61.
doi: 10.1107/S0907444910027915. Epub 2010 Oct 20.

Using neutron protein crystallography to understand enzyme mechanisms

Jenny P Glusker  1 , H L Carrell, Andrey Y Kovalevsky, Leif Hanson, S Zoe Fisher, Marat Mustyakimov, Sax Mason, Trevor Forsyth, Paul Langan
Affiliations

Using neutron protein crystallography to understand enzyme mechanisms

Jenny P Glusker et al. Acta Crystallogr D Biol Crystallogr. 2010 Nov.

Abstract

A description is given of the results of neutron diffraction studies of the structures of four different metal-ion complexes of deuterated D-xylose isomerase. These represent four stages in the progression of the biochemical catalytic action of this enzyme. Analyses of the structural changes observed between the various three-dimensional structures lead to some insight into the mechanism of action of this enzyme.

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Figures

Figure 1
Figure 1
A chemical diagram of the three-stage sugar-interconversion reaction catalyzed by XI, involving (i) ring-opening, (ii) isomerization and (iii) ring-closure steps.
Figure 2
Figure 2
The active site of native XI, showing polar residues binding the metal sites M1 and M2 and hydrophobic residues lining the sites. The catalytic water is labeled Wcat, while the two water molecules coordinated to M1 are labeled W2 and W3. The coordinates are from 2gve.
Figure 3
Figure 3
A chart of the neutron structures obtained for XI. Each complex represents an intermediate state on the sugar-interconversion reaction pathway.
Figure 4
Figure 4
The intermediate states along the sugar-interconversion reaction as observed in the neutron structures. (a) Cyclic substrate glucose bound, (b) linear substrate glucose bound after ring opening has occurred and (c) linear product xylulose bound after the ring-opening and isomerization steps have occurred. Metal coordination is represented by black dashed lines, while hydrogen bonding is represented by dotted brown lines. D atoms are colored green. Note the interaction of the catalytic OD− with the C1 methylene group of the sugar.
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References

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