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首页> 外文期刊>Biochemistry >Penetration of Short Fluorescence-Labeled Peptidesinto the Nucleus in HeLa Cells and in vitro Specific Interactionof the Peptides with Deoxyribooligonucleotides and DNA
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Penetration of Short Fluorescence-Labeled Peptidesinto the Nucleus in HeLa Cells and in vitro Specific Interactionof the Peptides with Deoxyribooligonucleotides and DNA

机译：短荧光标记肽在HeLa细胞中渗透入核中以及该肽与脱氧核糖寡核苷酸和DNA的体外特异性相互作用

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Marked fluorescence in cytoplasm, nucleus, and nucleolus was observed in HeLa cells after incubation with each of several fluorescein isothiocyanate-labeled peptides (epithalon, Ala-Glu-Asp-Gly; pinealon, Glu-Asp-Arg; testagen, Lys-Glu-Asp-Gly). This means that short biologically active peptides are able to penetrate into an animal cell and its nucleus and, in principle they may interact with various components of cytoplasm and nucleus including DNA and RNA. It was established that various initial (intact) peptides differently affect the fluorescence of the 5,6-carboxyfiuorescein-labeled deoxyribooligonucleotides and DNA-ethidium bromide complexes. The Stern-Volmer constants characterizing the degree of fluorescence quenching of various single- and double-stranded fluorescence-labeled deoxyribooligonucleotides with short peptides used were different depending on the peptide primary structures. This indicates the specific interaction between short biologically active peptides and nucleic acid structures. On binding to them, the peptides discriminate between different nucleotide sequences and recognize even their cytosine methylation status. Judging from corresponding constants of the fluorescence quenching, the epithalon, pinealon, and bronchogen (Ala-Glu-Aisp-Leu) bind preferentially with deoxyribooligonucleotides containing CNG sequence (CNG sites are targets for cytosine DNA methylation in eukary-otes). Epithalon, testagen, and pinealon seem to preferentially bind with CAG- but bronchogen with CTG-containing sequences. The site-specific interactions of peptides with DNA can control epigenetically the cell genetic functions, and they seem to play an important role in regulation of gene activity even at the earliest stages of life origin and in evolution.
机译：与几种荧光素异硫氰酸酯标记的肽（上皮，Ala-Glu-Asp-Gly;松果体，Glu-Asp-Arg;睾丸激素，Lys-Glu- Asp-Gly）。这意味着短的生物活性肽能够穿透动物细胞及其细胞核，并且原则上它们可以与细胞质和细胞核的各种成分（包括DNA和RNA）相互作用。已经确定，各种初始（完整）肽会不同地影响5，6-羧基荧光素标记的脱氧核糖寡核苷酸和DNA-溴化乙锭复合物的荧光。表征具有短肽的各种单链和双链荧光标记的脱氧核糖寡核苷酸的荧光猝灭程度的斯特恩-沃尔默常数根据肽的一级结构而不同。这表明短的生物活性肽和核酸结构之间的特异性相互作用。与它们结合后，这些肽可以区分不同的核苷酸序列，甚至可以识别其胞嘧啶甲基化状态。从荧光猝灭的相应常数来看，上皮，松果体和支气管原（Ala-Glu-Aisp-Leu）优先与含有CNG序列的脱氧核糖寡核苷酸结合（CNG位点是真核生物中胞嘧啶DNA甲基化的靶标）。 Epithalon，睾丸激素和松果体似乎优先结合CAG，但支气管原具有CTG的序列。肽与DNA的位点特异性相互作用可以表观遗传地控制细胞的遗传功能，即使在生命起源和进化的最早阶段，它们也似乎在调节基因活性中起着重要作用。

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