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

Elucidating the exact role of engineered CRABPII residues for the formation of a retinal protonated Schiff base.
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DOI:
10.1002/prot.22495
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发表时间:
2009-12
影响因子:
2.9
通讯作者:
Borhan, Babak
Borhan, Babak
中科院分区:
生物学4区
文献类型:
--
作者:
Vasileiou, Chrysoula;Wang, Wenjing;Jia, Xiaofei;Lee, Kin Sing Stephen;Watson, Camille T.;Geiger, James H.;Borhan, Babak

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细胞视黄酸结合蛋白II(CRABPII)已被重新设计,以特异性结合并与全反式视黄醇反应,形成质子化的席夫碱。该过程的每个步骤已被解剖,并且CRABPII结合位点内的四个残基(Lys 132,Tyr 134,Arg 111,Glu 121)已被鉴定为对亚胺形成和/或质子化至关重要。每个残基的确切作用已通过定点诱变和晶体学研究进行了检查。R132 K:L121 E-CRABPII双突变体的晶体结构表明工程Glu 121和天然Arg 111之间的直接相互作用,这对于席夫碱形成和质子化都是至关重要的。
Cellular Retinoic Acid Binding Protein II (CRABPII) has been re-engineered 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, 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 double mutant suggests a direct interaction between engineered Glu121 and the native Arg111, which is critical for both Schiff base formation and protonation.
DOI: 10.1111/j.1751-1097.1971.tb06192.x
发表时间: 1971-01-01
影响因子: 3.3
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