Microseeding – A Powerful Tool for Crystallizing Proteins Complexed with Hydrolyzable Substrates
Hydrolysis is an often-encountered obstacle in the crystallization of proteins complexed with their substrates. As the duration of the crystallization process, from nucleation to the growth of the crystal to its final size, commonly requires several weeks, non-enzymatic hydrolysis of an “unstab...
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| Format: | Article |
| Language: | English |
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MDPI AG
2008-07-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | http://www.mdpi.com/1422-0067/9/7/1131/ |
| _version_ | 1811304095687901184 |
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| author | Lutz Schmitt Erhard Bremer Sander H. J. Smits Christine Oswald |
| author_facet | Lutz Schmitt Erhard Bremer Sander H. J. Smits Christine Oswald |
| author_sort | Lutz Schmitt |
| collection | DOAJ |
| description | Hydrolysis is an often-encountered obstacle in the crystallization of proteins complexed with their substrates. As the duration of the crystallization process, from nucleation to the growth of the crystal to its final size, commonly requires several weeks, non-enzymatic hydrolysis of an “unstable†ligand occurs frequently. In cases where the crystallization conditions exhibit non neutral pH values this hydrolysis phenomenon may be even more pronounced. ChoX, the substrate binding protein of a choline ABC-importer, produced crystals with its substrate acetylcholine after one month. However, these crystals exhibited only choline, an acetylcholine hydrolysis product, in the binding site. To overcome this obstacle we devised a microseeding protocol leading to crystals of ChoX with bound acetylcholine within 24 hours. One drawback we encountered was the high twinning fraction of the crystals, possibly was due to the rapid crystal growth. |
| first_indexed | 2024-04-13T08:01:05Z |
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| id | doaj.art-1df7766345904e9585d84039310a05cf |
| institution | Directory Open Access Journal |
| issn | 1422-0067 |
| language | English |
| last_indexed | 2024-04-13T08:01:05Z |
| publishDate | 2008-07-01 |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-1df7766345904e9585d84039310a05cf2022-12-22T02:55:17ZengMDPI AGInternational Journal of Molecular Sciences1422-00672008-07-019711311141Microseeding – A Powerful Tool for Crystallizing Proteins Complexed with Hydrolyzable SubstratesLutz SchmittErhard BremerSander H. J. SmitsChristine OswaldHydrolysis is an often-encountered obstacle in the crystallization of proteins complexed with their substrates. As the duration of the crystallization process, from nucleation to the growth of the crystal to its final size, commonly requires several weeks, non-enzymatic hydrolysis of an “unstable†ligand occurs frequently. In cases where the crystallization conditions exhibit non neutral pH values this hydrolysis phenomenon may be even more pronounced. ChoX, the substrate binding protein of a choline ABC-importer, produced crystals with its substrate acetylcholine after one month. However, these crystals exhibited only choline, an acetylcholine hydrolysis product, in the binding site. To overcome this obstacle we devised a microseeding protocol leading to crystals of ChoX with bound acetylcholine within 24 hours. One drawback we encountered was the high twinning fraction of the crystals, possibly was due to the rapid crystal growth.http://www.mdpi.com/1422-0067/9/7/1131/crystal twinningacetylchline binding proteinco-crystallization |
| spellingShingle | Lutz Schmitt Erhard Bremer Sander H. J. Smits Christine Oswald Microseeding – A Powerful Tool for Crystallizing Proteins Complexed with Hydrolyzable Substrates International Journal of Molecular Sciences crystal twinning acetylchline binding protein co-crystallization |
| title | Microseeding – A Powerful Tool for Crystallizing Proteins Complexed with Hydrolyzable Substrates |
| title_full | Microseeding – A Powerful Tool for Crystallizing Proteins Complexed with Hydrolyzable Substrates |
| title_fullStr | Microseeding – A Powerful Tool for Crystallizing Proteins Complexed with Hydrolyzable Substrates |
| title_full_unstemmed | Microseeding – A Powerful Tool for Crystallizing Proteins Complexed with Hydrolyzable Substrates |
| title_short | Microseeding – A Powerful Tool for Crystallizing Proteins Complexed with Hydrolyzable Substrates |
| title_sort | microseeding a¢a€a a powerful tool for crystallizing proteins complexed with hydrolyzable substrates |
| topic | crystal twinning acetylchline binding protein co-crystallization |
| url | http://www.mdpi.com/1422-0067/9/7/1131/ |
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