Synthesis and characterization of a sequence-specific DNA-binding protein that contains ruthenium polypyridyl centers

被引：22
作者：
Lasey, RC
Banerji, SS
Ogawa, MY [1 ]
机构：
[1] Bowling Green State Univ, Dept Chem, Bowling Green, OH 43403 USA
[2] Bowling Green State Univ, Ctr Photochem Sci, Bowling Green, OH 43403 USA
[3] Med Coll Ohio, Dept Biochem & Mol Biol, Toledo, OH 43614 USA
来源：
INORGANICA CHIMICA ACTA | 2000年 / 300卷
基金：
美国国家卫生研究院;
关键词：
ruthenium; de novo protein; bZIP; GCN-4; DNA;
D O I：
10.1016/S0020-1693(99)00589-7
中图分类号：
O61 [无机化学];
学科分类号：
070301 ; 081704 ;
摘要：
A chimeric metallo-bZIP protein, (GBR-CC)Ru, was prepared that contains the native DNA-binding domain of the GCN-4 transcription factor (GBR), a synthetic alpha-helical coiled-coil dimerization site (CC), and a ruthenium polypyridyl complex (Ru) attached to a surface-exposed cysteine residue. The photophysical properties of the peptide-bound ruthenium complex include a long-lived emission that is shortened when the peptide is dissolved in air-saturated water. Electrophoretic mobility shift assays show that the chimeric metalloprotein also retains the essential DNA recognition properties of the native GCN-4 transcriptional activator. Peptide-DNA complexes are formed with the related AP1 and CRE sequences, but not the divergent Spl sequence. Peptide titration studies indicate that the affinities of (GBR-CC)Ru for the AP1 and CRE sites are comparable. Steady-state photolysis experiments show that (GBR-CC)Ru does not produce photoinduced DNA damage, probably due to the separation distance between the ruthenium sites and the DNA. bases. (C) 2000 Elsevier Science S.A. All rights reserved.
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收藏
页码：822 / 828
页数：7
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