Solving the phase problem for carbohydrate-binding proteins using selenium derivatives of their ligands: a case study involving the bacterial F17-G adhesin.

TitleSolving the phase problem for carbohydrate-binding proteins using selenium derivatives of their ligands: a case study involving the bacterial F17-G adhesin.
Publication TypeJournal Article
Year of Publication2003
AuthorsButs, L., R. Loris, E. De Genst, S. Oscarson, M. Lahmann, J. Messens, E. Brosens, L. Wyns, H. De Greve, and J. Bouckaert
JournalActa Crystallogr D Biol Crystallogr
Volume59
IssuePt 6
Pagination1012-5
Date Published2003 Jun
ISSN0907-4449
KeywordsAcetylglucosamine, Adhesins, Bacterial, Carrier Proteins, Crystallization, Escherichia coli Proteins, Hydrogen Bonding, Indicators and Reagents, Ligands, Models, Molecular, Protein Binding, Receptors, Cell Surface, Selenium Compounds
Abstract

The Escherichia coli adhesin F17-G is a carbohydrate-binding protein that allows the bacterium to attach to the intestinal epithelium of young ruminants. The structure of the 17 kDa lectin domain of F17-G was determined using the anomalous dispersion signal of a selenium-containing analogue of the monosaccharide ligand N-acetyl-d-glucosamine in which the anomeric oxygen was replaced by an Se atom. A three-wavelength MAD data set yielded good experimental phases to 2.6 A resolution. The structure was refined to 1.75 A resolution and was used to solve the structures of the ligand-free protein and the F17-G-N-acetyl-d-glucosamine complex. This selenium-carbohydrate phasing method could be of general use for determining the structures of carbohydrate-binding proteins.

Alternate JournalActa Crystallogr. D Biol. Crystallogr.
PubMed ID12777763
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