Title | Impact of natural variation in bacterial F17G adhesins on crystallization behaviour. |
Publication Type | Journal Article |
Year of Publication | 2005 |
Authors | Buts, L., A. Wellens, I. Van Molle, L. Wyns, R. Loris, M. Lahmann, S. Oscarson, H. De Greve, and J. Bouckaert |
Journal | Acta Crystallogr D Biol Crystallogr |
Volume | 61 |
Issue | Pt 8 |
Pagination | 1149-59 |
Date Published | 2005 Aug |
ISSN | 0907-4449 |
Keywords | Adhesins, Bacterial, Amino Acid Sequence, Crystallization, Crystallography, X-Ray, Disaccharides, Escherichia coli Proteins, Genetic Variation, Models, Molecular, Molecular Sequence Data, Protein Structure, Tertiary, Sequence Alignment |
Abstract | Since the introduction of structural genomics, the protein has been recognized as the most important variable in crystallization. Recent strategies to modify a protein to improve crystal quality have included rationally engineered point mutations, truncations, deletions and fusions. Five naturally occurring variants, differing in 1-18 amino acids, of the 177-residue lectin domain of the F17G fimbrial adhesin were expressed and purified in identical ways. For four out of the five variants crystals were obtained, mostly in non-isomorphous space groups, with diffraction limits ranging between 2.4 and 1.1 A resolution. A comparative analysis of the crystal-packing contacts revealed that the variable amino acids are often involved in lattice contacts and a single amino-acid substitution can suffice to radically change crystal packing. A statistical approach proved reliable to estimate the compatibilities of the variant sequences with the observed crystal forms. In conclusion, natural variation, universally present within prokaryotic species, is a valuable genetic resource that can be favourably employed to enhance the crystallization success rate with considerably less effort than other strategies. |
DOI | 10.1107/S0907444905017038 |
Alternate Journal | Acta Crystallogr. D Biol. Crystallogr. |
PubMed ID | 16041081 |
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