Action pattern of Fusarium moniliforme endopolygalacturonase towards pectin fragments: Comprehension and prediction

• Published in: Biochimica et biophysica acta Vol. 1749; no. 1; pp. 53 - 64
• Main Authors: André-Leroux, G., Tessier, D., Bonnin, E.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 20.05.2005; Elsevier
• Subjects: Action pattern; Amino Acid Sequence; Binding Sites; Chemical and Process Engineering; Computational Biology; Endopolygalacturonase; Engineering Sciences; Food engineering; Fusarium - enzymology; Homogalacturonan; Life Sciences; Models, Chemical; Molecular modeling; Molecular Sequence Data; Molecular Structure; Pectins - chemistry; Polygalacturonase - chemistry; Protein Conformation; Sequence Alignment; Subsite binding; Action pattern; Endopolygalacturonase; Subsite binding; Homogalacturonan; Molecular modeling
• ISSN: 1570-9639; 0006-3002; 1878-1454
• DOI: 10.1016/j.bbapap.2005.02.008

The structures of complexes of Fusarium moniliforme endopolygalacturonase (endoPG) with non-methylated or partly methylated homogalacturonan fragments were modeled to identify the residues involved in substrate binding and to correlate the cleavage pattern with the experimental productive modes. The conformational space of the complex was extensively explored and malto- to hexo-oligogalacturonates were modeled in the active cleft. To select the most highly probable productive complex for each oligomer between DP2 and 6, four energetic criteria were defined. Noteworthingly, the results were in accordance with the experimental results showing the mode of action of this enzyme towards un-methyl-esterified oligogalacturonates. Furthermore, the amino-acid residues involved in the binding were confirmed by similar studies performed on other endoPGs. Then, the oligomers were gradually methyl-esterified at one or more positions and similar docking experiments were carried out. Markedly, the docking energies were not significantly modified by the methyl-esterification of the substrate and it is likely that the methyl-esterification of the substrate does not alter the mode of action of the enzyme. Finally, 1D sequence and 3D structure of the endopolygalacturonase of Aspergillus niger II, known to be strictly non-tolerant to methylesters, were compared with the sequence and structure of the tolerant F. moniliforme endopolygalacturonase to get to a structural comprehension of the tolerant–or not–behaviour of endoPGs with methyl-esterified pectins.