Comparative Membrane Proteomics Reveals a Nonannotated E. coli Heat Shock Protein.
Comparative Membrane Proteomics Reveals a Nonannotated E. coli Heat Shock Protein.
复制标题
比较膜蛋白质组学揭示了一种未注释的大肠杆菌热休克蛋白。
DOI：
10.1021/acs.biochem.7b00864
复制
发表时间：
2018
期刊：
影响因子：
2.9
通讯作者：
Slavoff,SarahA
中科院分区：
文献类型：
--
作者：
Yuan,Peijia;D'Lima,NadiaG;Slavoff,SarahA
蛋白质组学和基因组学的最新进展使得在整个进化空间中发现了数千个先前未注释的小开放阅读框架（smORF）。此外，定量质谱法最近已被应用于分析调节smORF表达。然而，自下而上的蛋白质组学对膜蛋白仍然相对不敏感，这表明它们在之前的研究中可能被低估了。在这份报告中，我们增加了生化膜蛋白富集，我们以前开发的无标记定量蛋白质组学协议，揭示了一个从来没有以前确定的热休克蛋白inEscherichia coliK 12。这种推测的smORF编码的热休克蛋白，GndA，很可能是36-55个氨基酸的长度，并包含一个预测的跨膜螺旋。我们验证了热休克调节表达的thegndAsmORF，并证明GndA-GFP融合蛋白cofractionates与细胞膜。因此，定量膜蛋白质组学能够揭示可能在细菌应激反应中发挥作用的未注释的小蛋白质。
Recent advances in proteomics and genomics have enabled discovery of thousands of previously nonannotated small open reading frames (smORFs) in genomes across evolutionary space. Furthermore, quantitative mass spectrometry has recently been applied to analysis of regulated smORF expression. However, bottom-up proteomics has remained relatively insensitive to membrane proteins, suggesting they may have been underdetected in previous studies. In this report, we add biochemical membrane protein enrichment to our previously developed label-free quantitative proteomics protocol, revealing a never-before-identified heat shock protein inEscherichia coliK12. This putative smORF-encoded heat shock protein, GndA, is likely to be ∼36–55 amino acids in length and contains a predicted transmembrane helix. We validate heat shock-regulated expression of thegndAsmORF and demonstrate that a GndA-GFP fusion protein cofractionates with the cell membrane. Quantitative membrane proteomics therefore has the ability to reveal nonannotated small proteins that may play roles in bacterial stress responses.