Validation of Shewanella oneidensis MR-1 small proteins by AMT tag-based proteome analysis
Validation of Shewanella oneidensis MR-1 small proteins by AMT tag-based proteome analysis
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DOI：
10.1089/omi.2004.8.239
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发表时间：
2004-09-01
影响因子：
3.3
通讯作者：
Lipton, MS
中科院分区：
文献类型：
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作者：
Romine, MF;Elias, DA;Lipton, MS
使用准确质量和时间 (AMT) 标签质谱方法进行蛋白质鉴定的严格标准，我们在 Shewanella oneidensis MR-1 蛋白质的 172 个全局胰蛋白酶消化物中检测到了长度为 101 个氨基酸的 36 个蛋白质，其中包括 10 个被注释为假设蛋白质的蛋白质。映射到保守但功能未表征的蛋白质 SO4134 和 SO2787 的肽是这些样品中最常检测到的肽，而映射到假设的 SO2669 和 SO2063、保守的假设 SO0335 和 SO2176 以及 SlyX 蛋白 (SO1063) 的肽的观察频率与必需小蛋白（核糖体蛋白和翻译蛋白）的频率相似。因子 IF-1），表明它们可能具有类似重要的细胞功能。此外，还检测到了映射到 30 个基因的肽，这些基因预计编码移码、点突变或重新编码信号。在这 30 个基因中，在 13 个基因（SO0101、SO0419、SO0590、SO0738、SO1113、SO1211、SO3079、SO3130、SO3240、SO4231、SO4328、SO4422 和 SO4657）中检测到映射到内部终止密码子之外的位置的肽。虽然 SO0101 编码的全长甲酸脱氢酶的表达可以通过在内部终止密码子处掺入硒代半胱氨酸来解释，但其余基因中下游序列的翻译机制仍然未知。
Using stringent criteria for protein identification by accurate mass and time (AMT) tag mass spectrometric methodology, we detected 36 proteins of 101 amino acids in length, including 10 that were annotated as hypothetical proteins, in 172 global tryptic digests of Shewanella oneidensis MR-1 proteins. Peptides that map to the conserved, but functionally uncharacterized proteins SO4134 and SO2787, were the most frequently detected peptides in these samples, while those that map to hypotheticals SO2669 and SO2063, conserved hypotheticals SO0335 and SO2176, and the SlyX protein (SO1063) were observed at frequencies similar to those from essential small proteins (ribosomal proteins and translation. initiation factor IF-1), suggesting that they may function in similarly important cellular functions. In addition, peptides were detected that map to 30 genes predicted to encode frameshifts, point mutations, or recoding signals. Of these 30 genes, peptides that map to positions beyond internal stop codons were detected in 13 genes (SO0101, SO0419, SO0590, SO0738, SO1113, SO1211, SO3079, SO3130, SO3240, SO4231, SO4328, SO4422, and SO4657). While expression of the full-length formate dehydrogenase encoded by SO0101 can be explained by incorporation of selenocysteine at the internal stop codon, the mechanism of translating downstream sequences in the remaining genes remains unknown.