荧光探针与大肠杆菌苯丙氨酸转移核糖核酸X碱基的共价连接。

Covalent attachment of fluorescent probes to the X-base of Escherichia coli phenylalanine transfer ribonucleic acid.

作者信息

Schiller P W, Schechter A N

出版信息

Nucleic Acids Res. 1977 Jul;4(7):2161-7. doi: 10.1093/nar/4.7.2161.

DOI:10.1093/nar/4.7.2161

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC342555/

Abstract

tRNA PheE, coli was labeled with the N-hydroxysuccinimide esters of 1-dimethylaminonaphthalene-5-sulfonyl glycine and N-methylanthranilic acid through reaction with the amino acid moiety of its X-base, whereby yields of 66% and 24%, respectively, were obtained. The purified dimethylaminonaphthalene-sulfonate derivative could not be aminoacylated and was found to be a strong competitive inhibitor of phenylalanine-tRNA synthetase [Ki=8X10(-7) M]. The N-methylanthraniloyl derivative could be charged to an extent of 5% as compared to native tRNA Phe. The fluorescence emission spectra of the derivatives are indicative of a slightly hydrophobic environment for both fluorophores. The results suggest that the integrity of the polar amino acid group of the X-base is required for the maintenance of the biologically active conformation.

摘要

大肠杆菌苯丙氨酸tRNA（tRNA PheE, coli）通过与X碱基的氨基酸部分反应，用1-二甲基氨基萘-5-磺酰甘氨酸和N-甲基邻氨基苯甲酸的N-羟基琥珀酰亚胺酯进行标记，分别得到了66%和24%的产率。纯化后的二甲基氨基萘磺酸盐衍生物不能进行氨基酰化，并且被发现是苯丙氨酸-tRNA合成酶的一种强竞争性抑制剂[抑制常数Ki = 8×10⁻⁷ M]。与天然tRNA Phe相比，N-甲基邻氨基苯甲酰基衍生物的氨基酰化程度可达5%。衍生物的荧光发射光谱表明，两种荧光团所处的环境都略显疏水。结果表明，X碱基极性氨基酸基团的完整性对于维持生物活性构象是必需的。

相似文献

1

Covalent attachment of fluorescent probes to the X-base of Escherichia coli phenylalanine transfer ribonucleic acid.

Nucleic Acids Res. 1977 Jul;4(7):2161-7. doi: 10.1093/nar/4.7.2161.

2

[Chemical modification of the complex of tRNA Phe with phenylalanyl-tRNA synthetase from Escherichia coli].

Mol Biol (Mosk). 1979 Sep-Oct;13(5):994-1000.

3

[Modification of one tRNA recognition site of phenylalanyl-tRNA synthetase from E. coli MRE-600 with N-chlorambucilyl-phenylalanyl-tRNA].

Mol Biol (Mosk). 1978 Sep-Oct;12(5):1085-95.

4

[Chemical modification of phenylalanyl-tRNA synthetase and ribosomes of Escherichia coli with derivatives of tRNA-Phe carrying photoreactive groups on guanosine residues].

Mol Biol (Mosk). 1980 May-Jun;14(3):531-8.

5

Conservation in evolution for a small monomeric phenylalanyl-tRNA synthetase of the tRNA(Phe) recognition nucleotides and initial aminoacylation site.

Biochemistry. 1996 Jan 9;35(1):117-23. doi: 10.1021/bi9517998.

6

[Alkylation of tRNA-Phe free and bound to phenylalanyl-tRNA synthetase with 4-(N-2-chloroethyl-N-methylamino)benzylamine].

Mol Biol (Mosk). 1981 Jan-Feb;15(1):45-53.

7

[Specific modification of phenylalanine:tRNA-ligases of E. coli MRE-600 with N-chlorambucilyl-14c-phenylalanyl-tRNA].

Mol Biol (Mosk). 1975 Jul-Aug;9(4):509-15.

8

On the Phe-tRNA induced binding of fluorescent oligonucleotides to the ribosomal decoding site.

Nucleic Acids Res. 1977 Aug;4(8):2881-92. doi: 10.1093/nar/4.8.2881.

9

Kinetic analysis of an E.coli phenylalanine-tRNA synthetase mutant.

Nucleic Acids Res. 1988 Aug 11;16(15):7477-86. doi: 10.1093/nar/16.15.7477.

10

Characterisation of a new, fully active fluorescent derivative of E. coli tRNA Phe.

Nucleic Acids Res. 1980 Feb 25;8(4):827-43.

引用本文的文献

1

Detection of nucleic acid modifications by chemical reagents.

RNA Biol. 2017 Sep 2;14(9):1166-1174. doi: 10.1080/15476286.2016.1261788. Epub 2016 Nov 30.

2

Characterisation of a new, fully active fluorescent derivative of E. coli tRNA Phe.

Nucleic Acids Res. 1980 Feb 25;8(4):827-43.

3

Participation of X47-fluorescamine modified E. coli tRNAs in in vitro protein biosynthesis.

Nucleic Acids Res. 1978 Dec;5(12):4837-53. doi: 10.1093/nar/5.12.4837.

4

Ability of modified forms of phenylalanine tRNA to stimulate guanosine pentaphosphate synthesis by the stringent factor-ribosome complex of E. coli.

Nucleic Acids Res. 1978 Apr;5(4):1325-34. doi: 10.1093/nar/5.4.1325.

本文引用的文献

1

Replacement of Y base, dihydrouracil, and 7-methylguanine in tRNA by artificial odd bases.

FEBS Lett. 1971 Nov 1;18(2):214-218. doi: 10.1016/0014-5793(71)80447-7.

2

The synthesis of N-acetylphenylalanyl-sRNA.

Biochem Biophys Res Commun. 1966 Oct 5;25(1):17-22. doi: 10.1016/0006-291x(66)90633-4.

3

Fluorescence studies of nucleotides and polynucleotides. I. Formycin, 2-aminopurine riboside, 2,6-diaminopurine riboside, and their derivatives.

J Biol Chem. 1969 Mar 10;244(5):1228-37.

4

Studies of transfer RNA tertiary structure by singlet-singlet energy transfer.

Proc Natl Acad Sci U S A. 1970 Jan;65(1):39-46. doi: 10.1073/pnas.65.1.39.

5

Studies on polynucleotides. LXXV. Specificity of tRNA for codon recognition as studied by the ribosomal binding technique.

J Mol Biol. 1967 Oct 14;29(1):97-112. doi: 10.1016/0022-2836(67)90183-0.

6

3-(3-amino-3-carboxy-n-propyl)uridine. The structure of the nucleoside in Escherichia coli transfer ribonucleic acid that reacts with phenoxyacetoxysuccinimide.

Biochemistry. 1974 Jul 2;13(14):2932-7. doi: 10.1021/bi00711a024.

7

3-(3-Amino-3-carboxypropyl)uridine: a novel modified nucleoside isolated from Escherichia coli phenylalanine transfer ribonucleic acid.

Biochemistry. 1974 Jun 4;13(12):2620-5. doi: 10.1021/bi00709a023.

8

Activation of several tRNAs of Escherichia coli by the phenoxyacetyl derivative of N-hydroxysuccinimide.

Arch Biochem Biophys. 1974 Feb;160(2):631-42. doi: 10.1016/0003-9861(74)90440-8.

9

Patterns of base modification in tRNA.

Nat New Biol. 1973 Jul 4;244(131):18-20. doi: 10.1038/newbio244018a0.

10

Acylation of transfer ribonucleic acid with the N-hydroxysuccinimide ester of phenoxyacetic acid.

Biochemistry. 1972 Aug 29;11(18):3435-43. doi: 10.1021/bi00768a017.

文献AI研究员

20分钟写一篇综述，助力文献阅读效率提升50倍。

用中文搜PubMed

大模型驱动的PubMed中文搜索引擎

文档翻译

学术文献翻译模型，支持多种主流文档格式。