Cloning, Expression, Purification, Crystallization and Preliminary X-Ray Crystallographic Analysis of C-12 Hydroxylase EryK from Saccharopolyspora erythraea
| Autor: | Steven G. Kendrew, Giuliano Sciara, Adriana E. Miele, Carmelinda Savino, Beatrice Vallone |
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| Přispěvatelé: | Istituto di Biologia e Patologia Molecolari, CNR, and Dipartimento di Scienze Biochimiche, Università degli Studi di Roma 'La Sapienza' = Sapienza University [Rome], SGA Technologies Ltd, ESRF ID14-2, ID14-3-BESSY ID14.1, EU RII3-CT2004-506008 |
| Rok vydání: | 2008 |
| Předmět: |
cytochrome P450
Stereochemistry [SDV]Life Sciences [q-bio] 030303 biophysics Gene Expression Crystallography X-Ray Biochemistry law.invention 03 medical and health sciences Bacterial Proteins Cytochrome P-450 Enzyme System Structural Biology law Escherichia coli Molecular replacement Cloning Molecular Crystallization 030304 developmental biology 0303 health sciences biology C-12 hydroxylase Substrate (chemistry) Cytochrome P450 Space group General Medicine biology.organism_classification c-12 hydroxylase crystallization cytochrome p450 erythromycin x-ray x-ray crystallography Metabolic pathway Crystallography biology.protein Orthorhombic crystal system Saccharopolyspora erythraea Saccharopolyspora |
| Zdroj: | Protein and Peptide Letters Protein and Peptide Letters, Bentham Science Publishers, 2008, 15 (10), pp.1138-1141. ⟨10.2174/092986608786071201⟩ Protein and peptide letters 15 (2008): 1138–1141. doi:10.2174/092986608786071201 info:cnr-pdr/source/autori:Savino,C.; Miele,A.E.; Sciara,G.; Kendrew,S.G.; Vallone,B./titolo:Cloning, epression, purification, crystallization and preliminary X-ray crystallographic analyis of C-12 Hydroxylase EryK from Saccharoplyspora erythraea/doi:10.2174%2F092986608786071201/rivista:Protein and peptide letters (Print)/anno:2008/pagina_da:1138/pagina_a:1141/intervallo_pagine:1138–1141/volume:15 |
| ISSN: | 0929-8665 |
| Popis: | Erythromycin A is produced by Saccharopolyspora erythraea via a secondary metabolic pathway using several steps including glycosylations and hydroxylations of the first macrolide intermediate 6-deoxyerythronolide B. Erythromycin C-12 hydroxylase (CYP113A1), the P450 cytochrome active in the final stages of erythromycin biosynthesis, was cloned and expressed in E. coli. Different crystal forms were harvested from distinct crystallization conditions: two ligand-free forms, one substrate bound and two inhibitors-bound. All crystals belong either to the monoclinc P2(1) or to the orthorhombic P2(1)2(1)2(1) space groups, and exhibit diffraction limits ranging from 2.3 to 1.6 angstrom. The structures will be determined by molecular replacement. |
| Databáze: | OpenAIRE |
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