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8-甲氧基补骨脂素与转运RNA及富含5-氟尿嘧啶的转运RNA的光结合作用。

Photobinding of 8-methoxypsoralen to transfer RNA and 5-fluorouracil-enriched transfer RNA.

作者信息

Ou C N, Song P S

出版信息

Biochemistry. 1978 Mar 21;17(6):1054-9. doi: 10.1021/bi00599a018.

DOI:10.1021/bi00599a018
PMID:343811
Abstract

The photobinding of [3H]8MOP to tRNA upon irradiation at 365 nm in the absence of O2 was determined by gel filtration. The maximum photobinding was found to be ca. 4 mol of 8MOP er mol of tRNA and 5FU-tRNA, with an overall quantum yield of 2.3 X 10(-3). The photobinding kinetics for 8MOP-tRNA showed an apparent induction period or sigmoidal kinetic curve, indicating a specific initial photobinding site on tRNA which was identified as 4-thiouridine at position 8 from the 5'-end of Escherichia coli tRNA. Photobinding of 8MOP to 5FU-tRNA proceeded without an apparent induction period. 8MOP-tRNA and 8MOP-5FU-tRNA adducts were characterized by absorption, fluorescence, and CD spectroscopy. A modified procedure was also developed to analyze the nucleoside composition in modified 8MOP-tRNA and 8MOP-5FU-tRNA. The results showed that 8MOP photochemically added mainly to pyrimidine bases. The photobinding of 8MOP changed the conformation (secondary in particular) of tRNA and inhibited aminoacyl-tRNA synthetase activity.

摘要

在无氧条件下，通过凝胶过滤法测定了[3H]8-甲氧基补骨脂素（8MOP）在365nm光照下与转运核糖核酸（tRNA）的光结合情况。发现最大光结合量约为每摩尔tRNA和5-氟尿嘧啶转运核糖核酸（5FU-tRNA）含有4摩尔8MOP，总量子产率为2.3×10⁻³。8MOP-tRNA的光结合动力学呈现出明显的诱导期或S形动力学曲线，这表明tRNA上存在一个特定的初始光结合位点，该位点被确定为大肠杆菌tRNA 5'端第8位的4-硫尿苷。8MOP与5FU-tRNA的光结合过程没有明显的诱导期。通过吸收光谱、荧光光谱和圆二色光谱对8MOP-tRNA和8MOP-5FU-tRNA加合物进行了表征。还开发了一种改进的方法来分析修饰后的8MOP-tRNA和8MOP-5FU-tRNA中的核苷组成。结果表明，8MOP主要通过光化学作用加成到嘧啶碱基上。8MOP的光结合改变了tRNA的构象（特别是二级结构），并抑制了氨酰-tRNA合成酶的活性。

相似文献

1
Photobinding of 8-methoxypsoralen to transfer RNA and 5-fluorouracil-enriched transfer RNA.8-甲氧基补骨脂素与转运RNA及富含5-氟尿嘧啶的转运RNA的光结合作用。
Biochemistry. 1978 Mar 21;17(6):1054-9. doi: 10.1021/bi00599a018.
2
Position of aminoacylation of individual Escherichia coli and yeast tRNAs.单个大肠杆菌和酵母tRNA的氨酰化位置。
Proc Natl Acad Sci U S A. 1976 Feb;73(2):405-9. doi: 10.1073/pnas.73.2.405.
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The plant aminoacyl-tRNA synthetases. Effect of sodium chloride on tRNA aminoacylation and aminoacyl-tRNA decomposition catalysed by aminoacyl-tRNA synthetases from yellow lupin seeds.植物氨酰-tRNA合成酶。氯化钠对黄羽扇豆种子氨酰-tRNA合成酶催化的tRNA氨酰化及氨酰-tRNA分解的影响。
Acta Biochim Pol. 1977;24(2):163-70.
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Involvement of the anticodon region of Escherichia coli tRNAGln and tRNAGlu in the specific interaction with cognate aminoacyl-tRNA synthetase. Alteration of the 2-thiouridine derivatives located in the anticodon of the tRNAs by BrCN or sulfur deprivation.大肠杆菌谷氨酰胺转运RNA（tRNAGln）和谷氨酸转运RNA（tRNAGlu）反密码子区域在与同源氨酰-tRNA合成酶特异性相互作用中的作用。通过溴化氰（BrCN）或硫缺乏改变位于tRNA反密码子中的2-硫代尿苷衍生物。
Biochim Biophys Acta. 1974 May 31;349(3):328-38. doi: 10.1016/0005-2787(74)90120-8.
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Experimental evidence for kinetic proofreading in the aminoacylation of tRNA by synthetase.合成酶对tRNA进行氨酰化过程中动力学校正的实验证据。
Proc Natl Acad Sci U S A. 1977 Jun;74(6):2246-50. doi: 10.1073/pnas.74.6.2246.
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Covalent cross-linking of transfer ribonucleic acid to the ribosomal P site. Mechanism and site of reaction in transfer ribonucleic acid.转移核糖核酸与核糖体P位点的共价交联。转移核糖核酸中的反应机制和位点。
Biochemistry. 1979 Oct 2;18(20):4322-32. doi: 10.1021/bi00587a010.
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Amino acids are not all initially attached to the same position on transfer RNA molecules.氨基酸并非一开始都附着在转运RNA分子的同一位置上。
Proc Natl Acad Sci U S A. 1975 Aug;72(8):3044-8. doi: 10.1073/pnas.72.8.3044.
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Tyrosyl-tRNA synthetase from Escherichia coli. Stoichiometry of ligand binding and half-of-the-sites reactivity in aminoacylation.来自大肠杆菌的酪氨酰 - tRNA合成酶。氨基酰化过程中配体结合的化学计量和半位点反应活性
Biochemistry. 1975 Jul 29;14(15):3344-50. doi: 10.1021/bi00686a009.
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Recognition of tRNAs by aminoacyl-tRNA synthetases: Escherichia coli tRNAMet and E. coli methionyl-tRNA synthetase.氨酰-tRNA合成酶对tRNA的识别：大肠杆菌甲硫氨酸tRNA和大肠杆菌甲硫氨酰-tRNA合成酶
Fed Proc. 1984 Dec;43(15):2977-80.
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Mechanism of aminoacylation of tRNA. Proof of the aminoacyl adenylate pathway for the isoleucyl- and tyrosyl-tRNA synthetases from Escherichia coli K12.tRNA的氨酰化机制。来自大肠杆菌K12的异亮氨酰-tRNA合成酶和酪氨酰-tRNA合成酶的氨酰腺苷酸途径的证据。
Biochemistry. 1976 Feb 24;15(4):818-23. doi: 10.1021/bi00649a014.

引用本文的文献

1
Toxicological action and ecological importance of plant photosensitizers.植物光敏剂的毒理学作用和生态重要性。
J Chem Ecol. 1986 Apr;12(4):823-34. doi: 10.1007/BF01020254.
2
Identification of a site of psoralen crosslinking in E. coli 16S ribosomal RNA.大肠杆菌16S核糖体RNA中补骨脂素交联位点的鉴定。
Nucleic Acids Res. 1982 May 11;10(9):2839-49. doi: 10.1093/nar/10.9.2839.
3
Identification of the modified nucleotides produced by covalent photoaddition of hydroxymethyltrimethylpsoralen to RNA.鉴定羟甲基三甲基补骨脂素与RNA共价光加成产生的修饰核苷酸。
Nucleic Acids Res. 1981 May 11;9(9):2207-22. doi: 10.1093/nar/9.9.2207.
4
Spectroscopic properties of ethidium monoazide: a fluorescent photoaffinity label for nucleic acids.单叠氮溴化乙锭的光谱性质：一种用于核酸的荧光光亲和标记物。
Nucleic Acids Res. 1978 Dec;5(12):4891-903. doi: 10.1093/nar/5.12.4891.