Comprehensive analysis of phosphorylated proteins of Escherichia coli ribosomes.
Comprehensive analysis of phosphorylated proteins of Escherichia coli ribosomes.
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DOI：
10.1021/pr900042e
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发表时间：
2009-07
影响因子：
4.4
通讯作者：
Koc EC
中科院分区：
文献类型：
--
作者：
Soung GY;Miller JL;Koc H;Koc EC
细菌核糖体蛋白的磷酸化已经知道了几十年;然而，关于核糖体蛋白中磷酸化位点的具体位置以及它们在蛋白质合成中可能发挥的作用的信息仍然非常有限。在本研究中，我们已经定位了24个E。大肠杆菌核糖体蛋白的串联质谱分析。通过磷酸化特异性可视化技术、ProQ染色和磷酸化-Ser、Thr和Tyr抗体，或通过配备有检测磷酸部分的添加和损失的能力的质谱法，实现磷酸化的特异性检测。通过固定化金属亲和和/或强阳离子交换色谱的富集被用于提高低丰度磷酸肽的检测的成功率。我们发现小亚基（30 S）蛋白S3、S4、S5、S7、S11、S12、S13、S18和S21以及大亚基（50 S）蛋白L1、L2、L3、L5、L 6、L7/L12、L13、L14、L16、L18、L19、L21、L22、L28、L31在一个或多个残基处被磷酸化。通过对核糖体三维晶体结构模型中磷酸化位点的仔细评估和先前对E. coli核糖体蛋白。
Phosphorylation of bacterial ribosomal proteins has been known for decades; however, there is still very limited information available on specific locations of the phosphorylation sites in ribosomal proteins and the role they might play in protein synthesis. In this study, we have mapped the specific phosphorylation sites in twenty-four E. coli ribosomal proteins by tandem mass spectrometry. Specific detection of phosphorylation was achieved by either phosphorylation specific visualization techniques, ProQ staining and antibodies for phospho-Ser, Thr, and Tyr, or by mass spectrometry equipped with a capability to detect addition and the loss of the phosphate moiety. Enrichment by immobilized metal affinity and/or strong cation exchange chromatography was used to improve the success of detection of the low abundance phosphopeptides. We found the small subunit (30S) proteins S3, S4, S5, S7, S11, S12, S13, S18, and S21 and the large subunit (50S) proteins L1, L2, L3, L5, L6, L7/L12, L13, L14, L16, L18, L19, L21, L22, L28, L31 to be phosphorylated at one or more residues. Potential roles for each specific site in ribosome function were deduced through careful evaluation of the given site of the phosphorylation in 3D-crystal structure models of ribosomes and the previous mutational studies of E. coli ribosomal proteins.
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DOI：
10.1073/pnas.0607541103
发表时间：
2006-10-24
影响因子：
11.1
作者：
Berk, Veysel;Zhang, Wen;Cate, Jamie H. Doudna
通讯作者：
Cate, Jamie H. Doudna
影响因子：
64.8
作者：
Ban, N;Nissen, P;Steitz, TA
通讯作者：
Steitz, TA
影响因子：
64.8
作者：
FRANK, J;ZHU, J;AGRAWAL, RK
通讯作者：
AGRAWAL, RK
DOI：
10.1038/10695
发表时间：
1999-07-01
期刊：
NATURE STRUCTURAL BIOLOGY
影响因子：
--
作者：
Agrawal, RK;Heagle, AB;Frank, J
通讯作者：
Frank, J
影响因子：
3.5
作者：
BRANDT, R;GUALERZI, CO
通讯作者：
GUALERZI, CO