title: Crystallization and preliminary crystallographic analysis of the type IIL restriction enzyme MmeI in complex with DNA.
abstract: 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 Å, α = 72.79, β = 89.12, γ = 71.68°, and diffracted to 2.6 Å 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.
authors.affiliation: Department of Structural and Chemical Biology, Mount Sinai School of Medicine, Box 1677, New York, NY 10029-6574, USA.
authors.forename: Scott J
authors.initials: SJ
authors.lastname: Callahan
authors.forename: Richard D
authors.initials: RD
authors.lastname: Morgan
authors.forename: Rinku
authors.initials: R
authors.lastname: Jain
authors.forename: Sharon A
authors.initials: SA
authors.lastname: Townson
authors.forename: Geoffrey G
authors.initials: GG
authors.lastname: Wilson
authors.forename: Richard J
authors.initials: RJ
authors.lastname: Roberts
authors.forename: Aneel K
authors.initials: AK
authors.lastname: Aggarwal
mesh_terms: D003460:Crystallization; D018360:Crystallography, X-Ray; D004247:DNA / Q000737:chemistry* / Q000378:metabolism; D015252:Deoxyribonucleases, Type II Site-Specific / Q000737:chemistry* / Q000378:metabolism; D011485:Protein Binding
chemical_list: D004247:DNA; C049760:endodeoxyribonuclease MmeI; D015252:Deoxyribonucleases, Type II Site-Specific
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