Protein Design: Reengineering Cellular Retinoic Acid Binding Protein Ii into a Rhodopsin Protein Mimic.
From: Department of Chemistry, Michigan State University, East Lansing, Michigan 48824, USA.
Journal of the American Chemical Society
- Publish Date: May 2007
- ISSN: 0002-7863
- Volume: 129
- Issue: 19
- Pages: 6140-8
- Medium: Print
- Language: English
- Citation (JAMA): Vasileiou Chrysoula, Vaezeslami Soheila, Crist Rachael M, et al. Protein Design: Reengineering Cellular Retinoic Acid Binding Protein Ii into a Rhodopsin Protein Mimic.. J. Am. Chem. Soc. May 2007;129:6140-8
Abstract
Rational redesign of the binding pocket of Cellular Retinoic Acid Binding Protein II (CRABPII) has provided a mutant that can bind retinal as a protonated Schiff base, mimicking the binding observed in rhodopsin. The reengineering was accomplished through a series of choreographed manipulations to ultimately orient the reactive species (the epsilon-amino group of Lys132 and the carbonyl of retinal) in the proper geometry for imine formation. The guiding principle was to achieve the appropriate Bürgi-Dunitz trajectory for the reaction to ensue. Through crystallographic analysis of protein mutants incapable of forming the requisite Schiff base, a highly ordered water molecule was identified as a key culprit in orienting retinal in a nonconstructive manner. Removal of the ordered water, along with placing reinforcing mutations to favor the desired orientation of retinal, led to a triple mutant CRABPII protein capable of nanomolar binding of retinal as a protonated Schiff base. The high-resolution crystal structure of all-trans-retinal bound to the CRABPII triple mutant (1.2 A resolution) unequivocally illustrates the imine formed between retinal and the protein.
Mesh Headings (Keywords): Binding Sites, Crystallography, X-Ray, Models, Chemical, Models, Molecular, Mutation, Protein Binding, Protein Engineering, Protein Folding, Protein Structure, Secondary, Receptors, Retinoic Acid, Retinaldehyde, Rhodopsin, Schiff Bases
Check for Full Text / PubMed Unique Identifier (PMID): 17447762
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