Expression, crystallization and preliminary X-ray diffraction studies on the complete choline-binding domain of the major pneumococcal autolysin
| Autor: | Álvaro Ramón, Carlos Fernández-Tornero, Guillermo Giménez-Gallego, Antonio A. Romero, Concha Fernández-Cabrera |
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| Rok vydání: | 2002 |
| Předmět: |
Models
Molecular Protein Conformation Crystallography X-Ray medicine.disease_cause Choline law.invention Protein structure Structural Biology law medicine Binding site N-acetylmuramoyl-L-alanine amidase Escherichia coli Choline binding Binding Sites Strain (chemistry) Chemistry Autolysin N-Acetylmuramoyl-L-alanine Amidase General Medicine Recombinant Proteins Protein Structure Tertiary Crystallography Streptococcus pneumoniae Recombinant DNA Crystallization |
| Zdroj: | Acta Crystallographica Section D Biological Crystallography. 58:556-558 |
| ISSN: | 0907-4449 |
| DOI: | 10.1107/s0907444902000537 |
| Popis: | The major pneumococcal autolysin (LytA), a virulence factor of this bacterium, is composed of an amino-terminal catalytic domain plus a carboxyl-terminal choline-binding domain (ChBD). This C-terminal domain, responsible for anchorage to the cell wall, is a tandem of six imperfect 20-residue repeats whose precise ends have been difficult to establish by sequence methods. The reported crystal structure of a shortened C-terminal fragment of the protein suggested that it might contain an additional repeat and thus an additional choline-binding site (ChBS). The complete recombinant choline-binding domain of LytA has now been overexpressed in soluble form using a secreting Escherichia coli strain which facilitates purification with a higher yield. It has been crystallized at room temperature using MPD as the main precipitant. The crystals belong to space group P2(1) and diffract to beyond 3.2 A resolution on a synchrotron-radiation source. The molecular-replacement solution indicates that a new ChBS which fits the topology of the solenoid structure is formed in the N-terminal region. |
| Databáze: | OpenAIRE |
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