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537 a.a.
Key reference
DOI no: 10.1002/prot.21700 Proteins 71:552-564 (2008) PubMed id: 17963237
An alternate sucrose binding mode in the E203Q Arabidopsis invertase mutant: an X-ray crystallography and docking study. J.Mátrai, W.Lammens, A.Jonckheer, K.Le Roy, A.Rabijns, W.Van den Ende, M.De Maeyer. ABSTRACT
In the present study, we report on the X-ray crystallographic structure of a GH32 invertase mutant, (i.e., the Arabidopsis thaliana cell-wall invertase 1-E203Q, AtcwINV1-mutant) in complex with sucrose. This structure was solved to reveal the features of sugar binding in the catalytic pocket. However, as demonstrated by the X-ray structure the sugar binding and the catalytic pocket arrangement is significantly altered as compared with what was expected based on previous X-ray structures on GH-J clan enzymes. We performed a series of docking and molecular dynamics simulations on various derivatives of AtcwINV1 to reveal the reasons behind this modified sugar binding. Our results demonstrate that the E203Q mutation introduced into the catalytic pocket triggers conformational changes that alter the wild type substrate binding. In addition, this study also reveals the putative productive sucrose binding modus in the wild type enzyme.
Selected figure(s)
Figure 1.
Figure 1. (a) AtcwINV1 and (b) AtcwINV1-mutant-sucrose structures. The N-acetylglucosamine residues and glycosyltation sites are indicated by orange color. (c) and (d) panels show a zoom in into the catalytic pocket of the AtcwINV1 and AtcwINV1-mutant enzymes, respectively.Figure 2.
Figure 2. Representation of the sucroses/raffinose as they appear in the superimposed X-ray structures. Color codes are as follows: green: FEH IIa/sucrose (the FM modus), red: levansucrase (E342A)/sucrose (the LSM modus), orange:-fructosidase/raffinose, blue: invertase(E203Q)/sucrose (the MIM modus).
The above figures are reprinted by permission from John Wiley & Sons, Inc.: Proteins (2008, 71, 552-564) copyright 2008. Figures were selected by an automated process.
Literature references that cite this PDB file's key reference
PubMed id Reference
19936386 A.Homann, and J.Seibel (2009).
Chemo-enzymatic synthesis and functional analysis of natural and modified glycostructures.Nat Prod Rep, 26, 1555-1571.
19474088 T.Welham, J.Pike, I.Horst, E.Flemetakis, P.Katinakis, T.Kaneko, S.Sato, S.Tabata, J.Perry, M.Parniske, and T.L.Wang (2009).
A cytosolic invertase is required for normal growth and cell development in the model legume, Lotus japonicus.J Exp Bot, 60, 3353-3365.
19129163 W.Lammens, K.Le Roy, L.Schroeven, A.Van Laere, A.Rabijns, and W.Van den Ende (2009).
Structural insights into glycoside hydrolase family 32 and 68 enzymes: functional implications.J Exp Bot, 60, 727-740.
18802643 S.K.Seo, and A.Wei (2008).
Probing osmotic effects on invertase with L-(-)-sucrose.Org Biomol Chem, 6, 3362-3365. The most recent references are shown first. Citation data come partly from CiteXplore and partly from an automated harvesting procedure. Note that this is likely to be only a partial list as not all journals are covered by either method. However, we are continually building up the citation data so more and more references will be included with time.
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