title: 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.
authors: Department of Chemistry, Michigan State University, East Lansing, Michigan 48824, USA.
authors: Chrysoula
authors: C
authors: Vasileiou
authors: Wenjing
authors: W
authors: Wang
authors: Xiaofei
authors: X
authors: Jia
authors: Kin Sing Stephen
authors: KS
authors: Lee
authors: Camille T
authors: CT
authors: Watson
authors: James H
authors: JH
authors: Geiger
authors: Babak
authors: B
authors: Borhan
mesh_terms: D019943:Amino Acid Substitution; D001120:Arginine / Q000737:chemistry; D001483:Base Sequence; D001665:Binding Sites; D018360:Crystallography, X-Ray; D017931:DNA Primers / Q000235:genetics; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D015202:Protein Engineering; D011522:Protons; D018168:Receptors, Retinoic Acid / Q000737:chemistry* / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D012172:Retinaldehyde / Q000737:chemistry; D012545:Schiff Bases / Q000737:chemistry; D013050:Spectrometry, Fluorescence; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D055672:Static Electricity; D014364:Tryptophan / Q000737:chemistry; D014443:Tyrosine / Q000737:chemistry
chemical_list: D017931:DNA Primers; D011522:Protons; D018168:Receptors, Retinoic Acid; D011994:Recombinant Proteins; D012545:Schiff Bases; C082928:retinoic acid binding protein II, cellular; D014443:Tyrosine; D014364:Tryptophan; D001120:Arginine; D012172:Retinaldehyde