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用AUG底物类似物对核糖体解码位点进行亲和标记。

Affinity labeling of the ribosomal decoding site with an AUG-substrate analog.

作者信息

Pongs O, Lanka E

出版信息

Proc Natl Acad Sci U S A. 1975 Apr;72(4):1505-9. doi: 10.1073/pnas.72.4.1505.

DOI:10.1073/pnas.72.4.1505
PMID:1093183
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC432565/
Abstract

The trinucleotide AUG was condensed at the 5'-end with N-bromoacetyl-p-aminophenylphosphate. This bromoactylated AUG analog reacted irreversibly with the mRNA binding site of Escherichia coli 70S ribosomes. After reaction of 70S ribosomes with the AUG analog, labeled 30S subunits could be isolated that were programmed for initiation-factor-dependent binding of fMet-tRNAfMet. This shows that this AUG-affinity label reacted in the decoding site for fMet-tRNAfMet. By combination of sodium dodecyl sulfate-, Sarkosyl-, and ureapolyacrylamide gel electrophoresis the AUG-affinity label was found to be irreversibly bound to ribosomal proteins S4, the ram gene product, and S18.

摘要

三核苷酸AUG在5'-末端与N-溴乙酰基对氨基苯磷酸缩合。这种溴乙酰化的AUG类似物与大肠杆菌70S核糖体的mRNA结合位点发生不可逆反应。70S核糖体与AUG类似物反应后，可以分离出标记的30S亚基，这些亚基被编程用于起始因子依赖的fMet-tRNAfMet结合。这表明这种AUG亲和标记在fMet-tRNAfMet的解码位点发生反应。通过十二烷基硫酸钠、 Sarkosyl和脲聚丙烯酰胺凝胶电泳的组合，发现AUG亲和标记不可逆地结合到核糖体蛋白S4（ram基因产物）和S18上。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8f12/432565/88f71ff256f3/pnas00047-0294-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8f12/432565/edb002bf0b93/pnas00047-0293-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8f12/432565/d935d2b64918/pnas00047-0294-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8f12/432565/88f71ff256f3/pnas00047-0294-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8f12/432565/edb002bf0b93/pnas00047-0293-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8f12/432565/d935d2b64918/pnas00047-0294-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8f12/432565/88f71ff256f3/pnas00047-0294-b.jpg

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引用本文的文献

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3
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本文引用的文献

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Role of guanosine 5'-triphosphate in the initiation of Peptide synthesis, I. Synthesis of formylmethionyl-puromycin.鸟苷 5'-三磷酸在肽合成起始中的作用，I. 甲酰甲硫氨酰-嘌呤霉素的合成
Proc Natl Acad Sci U S A. 1967 Mar;57(3):759-66. doi: 10.1073/pnas.57.3.759.
2
The dependence of cell-free protein synthesis in E. coli upon naturally occurring or synthetic polyribonucleotides.大肠杆菌中无细胞蛋白质合成对天然存在或合成的多聚核糖核苷酸的依赖性。
Proc Natl Acad Sci U S A. 1961 Oct 15;47(10):1588-602. doi: 10.1073/pnas.47.10.1588.
3
Studies on polynucleotides. III. Enzymic degradation; substrate specificity and properties of snake venom phosphodiesterase.
Nucleic Acids Res. 1976 Jul;3(7):1625-33. doi: 10.1093/nar/3.7.1625.
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J Biol Chem. 1959 Aug;234(8):2105-13.
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The relationship between poly A polymerase and the ribosomes.
Biochemistry. 1966 Nov;5(11):3676-84. doi: 10.1021/bi00875a042.
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Role of 5S RNA in the functions of 50S ribosomal subunits.5S RNA在50S核糖体亚基功能中的作用。
Proc Natl Acad Sci U S A. 1971 Dec;68(12):2932-6. doi: 10.1073/pnas.68.12.2932.
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Ribosomal discrimination of tRNAs.核糖体对转运RNA的识别
Nat New Biol. 1971 Dec 29;234(52):261-4. doi: 10.1038/newbio234261a0.
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Alteration of a 30S ribosomal protein accompanying the ram mutation in Escherichia coli.伴随大肠杆菌中ram突变的30S核糖体蛋白的改变。
Proc Natl Acad Sci U S A. 1971 Sep;68(9):2263-7. doi: 10.1073/pnas.68.9.2263.
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