Selection of Peptides Targeting Helix 31 of Bacterial 16S Ribosomal RNA by Screening M13 Phage-Display Libraries
Selection of Peptides Targeting Helix 31 of Bacterial 16S Ribosomal RNA by Screening M13 Phage-Display Libraries
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DOI：
10.3390/molecules16021211
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发表时间：
2011-02-01
期刊：
影响因子：
4.6
通讯作者：
Chow, Christine S.
中科院分区：
文献类型：
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作者：
Lamichhane, Tek N.;Abeydeera, N. Dinuka;Chow, Christine S.
核糖体RNA是核糖体的催化部分，在翻译过程中经历各种构象变化。核糖体RNA的结构变化可以通过修饰的核苷酸的存在来促进。细菌16 S核糖体RNA的H31含有两个修饰的核苷酸，m（2）G966和m（5）C967，它们在细菌中高度保守，尽管在该区域的修饰程度和性质在真核生物中不同。螺旋31和P-位点tRNA、起始因子和核糖体蛋白之间的接触突出了该区域在翻译中的重要性。在这项工作中，一个七肽M13噬菌体展示库筛选的配体，目标野生型，天然修饰的细菌螺旋31。几轮筛选后，发现几种肽，包括TYLPWPA、CVRPFAL、TLWDLIP、FVRPFPL、ATPLWLK和DIRTQRE是普遍存在的。在生物物理测定中，包括表面等离子体共振（SPR），几种肽对修饰的螺旋31表现出中等亲和力（在高nM至低μ M范围内），并且还显示结合30 S核糖体亚基。这些肽还在无细胞翻译测定中抑制蛋白质合成。
Ribosomal RNA is the catalytic portion of ribosomes, and undergoes a variety of conformational changes during translation. Structural changes in ribosomal RNA can be facilitated by the presence of modified nucleotides. Helix 31 of bacterial 16S ribosomal RNA harbors two modified nucleotides, m(2)G966 and m(5)C967, that are highly conserved among bacteria, though the degree and nature of the modifications in this region are different in eukaryotes. Contacts between helix 31 and the P-site tRNA, initiation factors, and ribosomal proteins highlight the importance of this region in translation. In this work, a heptapeptide M13 phage-display library was screened for ligands that target the wild-type, naturally modified bacterial helix 31. Several peptides, including TYLPWPA, CVRPFAL, TLWDLIP, FVRPFPL, ATPLWLK, and DIRTQRE, were found to be prevalent after several rounds of screening. Several of the peptides exhibited moderate affinity (in the high nM to low mu M range) to modified helix 31 in biophysical assays, including surface plasmon resonance (SPR), and were also shown to bind 30S ribosomal subunits. These peptides also inhibited protein synthesis in cell-free translation assays.