Crystallization and preliminary crystallographic analysis of the type IIL restriction enzyme MmeI in complex with DNA
| Autor: | Geoffrey G. Wilson, Richard D. Morgan, Rinku Jain, Richard J. Roberts, Aneel K. Aggarwal, Scott J. Callahan, Sharon A. Townson |
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| Rok vydání: | 2011 |
| Předmět: |
chemistry.chemical_classification
Biophysics Substrate (chemistry) Plasma protein binding DNA Biology Condensed Matter Physics Crystallography X-Ray Biochemistry Amino acid Restriction enzyme chemistry.chemical_compound Crystallography Enzyme chemistry Structural Biology Crystallization Communications Genetics Deoxyribonucleases Crystallization Deoxyribonucleases Type II Site-Specific Peptide sequence Protein Binding |
| Zdroj: | Acta crystallographica. Section F, Structural biology and crystallization communications. 67(Pt 10) |
| ISSN: | 1744-3091 |
| Popis: | Type IIL restriction enzymes have rejuvenated the search for user-specified DNA binding and cutting. By aligning and contrasting the highly comparable amino-acid sequences yet diverse recognition specificities across the family of enzymes, amino acids involved in DNA binding have been identified and mutated to produce alternative binding specificities. To date, the specificity of MmeI (a type IIL restriction enzyme) has successfully been altered at positions 3, 4 and 6 of the asymmetric TCCRAC (where R is a purine) DNA-recognition sequence. To further understand the structural basis of MmeI DNA-binding specificity, the enzyme has been crystallized in complex with its DNA substrate. The crystal belonged to space group P1, with unit-cell parameters a = 61.73, b = 94.96, c = 161.24 A, α = 72.79, β = 89.12, γ = 71.68°, and diffracted to 2.6 A resolution when exposed to synchrotron radiation. The structure promises to reveal the basis of MmeI DNA-binding specificity and will complement efforts to create enzymes with novel specificities. |
| Databáze: | OpenAIRE |
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