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Elucidating the exact role of engineered CRABPII residues for the formation of a retinal protonated Schiff base.

ABSTRACT: Cellular Retinoic Acid Binding Protein II (CRABPII) has been reengineered to specifically bind and react with all-trans-retinal to form a protonated Schiff base. Each step of this process has been dissected and four residues (Lys132, Tyr134, Arg111, and Glu121) within the CRABPII binding site have been identified as crucial for imine formation and/or protonation. The precise role of each residue has been examined through site directed mutagenesis and crystallographic studies. The crystal structure of the R132K:L121E-CRABPII (PDB-3I17) double mutant suggests a direct interaction between engineered Glu121 and the native Arg111, which is critical for both Schiff base formation and protonation.

SUBMITTER: Vasileiou C

PROVIDER: S-EPMC4306427 | biostudies-literature | 2009 Dec

REPOSITORIES: biostudies-literature

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Elucidating the exact role of engineered CRABPII residues for the formation of a retinal protonated Schiff base.

Vasileiou Chrysoula C Wang Wenjing W Jia Xiaofei X Lee Kin Sing Stephen KS Watson Camille T CT Geiger James H JH Borhan Babak B

Proteins 20091201 4

Cellular Retinoic Acid Binding Protein II (CRABPII) has been reengineered to specifically bind and react with all-trans-retinal to form a protonated Schiff base. Each step of this process has been dissected and four residues (Lys132, Tyr134, Arg111, and Glu121) within the CRABPII binding site have been identified as crucial for imine formation and/or protonation. The precise role of each residue has been examined through site directed mutagenesis and crystallographic studies. The crystal structu ...[more]

PMID: 19603486

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