Structural Analysis of Cytochrome P450 105N1 Involved in the Biosynthesis of the Zincophore, Coelibactin
Abstract
:1. Introduction
2. Results and Discussion
2.1. The Role of the Coelibactin Operon in Zinc Chelation and Bioinformatics Analysis
2.2. Bioinformatic Analysis of CYP105N1
2.3. CYP105N1 Expression, Purification and Spectral Analysis
2.4. Features of the Crystal Structure of CYP105N1
2.4.1. The Overall Structure of CYP105N1
2.4.2. The Heme-Binding Pocket
2.4.3. Comparison of CYP105N1 with Other CYP105s
2.5. Conjecture on the Enzymatic Role(s) of CYP105N1 in Production of an Active Siderophore
3. Experimental Section
3.1. Cloning, Heterologous Expression and Purification of Streptomyces coelicolor A3(2) CYP105N1
3.2. Crystallization, Data collection and Structure Determination of CYP105N1
3.3. General Methods
4. Conclusions
Acknowledgments
References
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Data Collection Statistics | Substrate-free |
---|---|
Space group | P61 |
Unit Cell (Å) | a = b = 134.537 c = 230.58 |
Molecules/asymmetric unit | 4 |
Data resolution (Å) | 2.9 |
Redundancya | 16.3 (9.9) |
Completeness%a | 99.8 (99.8) |
I/σ(I)a | 26.8 (6.1) |
Rmerge%a | 7.1 (54.2) |
Refinement statistics | |
No. of reflections used in refinement | 51760 |
No. of water molecules | 100 |
Protein atoms | 12366 |
Heme atoms | 172 |
Ligand atoms | 0 |
Rwork% | 28.21 |
Rfree% | 30.05 |
Rmsdb in bond lengths (Å) | 0.009 |
Rmsd in bond angles (°) | 1.2 |
Ramachandran statistics | |
Favored regions (%) | 94.9 |
Allowed regions (%) | 3.2 |
Outerliers (%) | 1.9 |
© 2012 by the authors; licensee Molecular Diversity Preservation International, Basel, Switzerland. This article is an open-access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/3.0/).
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Zhao, B.; Moody, S.C.; Hider, R.C.; Lei, L.; Kelly, S.L.; Waterman, M.R.; Lamb, D.C. Structural Analysis of Cytochrome P450 105N1 Involved in the Biosynthesis of the Zincophore, Coelibactin. Int. J. Mol. Sci. 2012, 13, 8500-8513. https://doi.org/10.3390/ijms13078500
Zhao B, Moody SC, Hider RC, Lei L, Kelly SL, Waterman MR, Lamb DC. Structural Analysis of Cytochrome P450 105N1 Involved in the Biosynthesis of the Zincophore, Coelibactin. International Journal of Molecular Sciences. 2012; 13(7):8500-8513. https://doi.org/10.3390/ijms13078500
Chicago/Turabian StyleZhao, Bin, Suzy C. Moody, Robert C. Hider, Li Lei, Steven L. Kelly, Michael R. Waterman, and David C. Lamb. 2012. "Structural Analysis of Cytochrome P450 105N1 Involved in the Biosynthesis of the Zincophore, Coelibactin" International Journal of Molecular Sciences 13, no. 7: 8500-8513. https://doi.org/10.3390/ijms13078500