High throughput discovery of protein variants using proteomics informed by transcriptomics.

High throughput discovery of protein variants using proteomics informed by transcriptomics.
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DOI:
10.1093/nar/gky295
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发表时间:
2018-06-01
影响因子:
14.9
通讯作者:
Bessant C
Bessant C
中科院分区:
生物学2区
文献类型:
--
作者:
Saha S;Matthews DA;Bessant C

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转录组学(PIT)提供的蛋白质组学,其中针对从头组装转录本衍生的开放阅读框架搜索蛋白质组MS/MS光谱,可以揭示以前未知的翻译基因组元件(TGE)。然而,确定哪些TGE是真正的新的,哪些是已知蛋白质的变体,哪些只是不良序列组装的假象,是具有挑战性的。我们已经设计并实现了一个自动化的解决方案,通过比较参考蛋白质组序列来分类推定的TGE。这允许大规模鉴定序列多态性、剪接异构体和新的TGE,其由变体特异性肽证据的存在或不存在支持。与以前报道的方法不同,我们的方法不需要已知变体的目录,使其更适用于非模式生物。该方法在人类PIT数据上进行了验证,然后应用于小家鼠,狐蝠和埃及伊蚊。新的发现包括60种人类蛋白质亚型,32 392个多态性在P. alecto,和TGE与非甲硫氨酸起始位点,包括酪氨酸。
Proteomics informed by transcriptomics (PIT), in which proteomic MS/MS spectra are searched against open reading frames derived from de novo assembled transcripts, can reveal previously unknown translated genomic elements (TGEs). However, determining which TGEs are truly novel, which are variants of known proteins, and which are simply artefacts of poor sequence assembly, is challenging. We have designed and implemented an automated solution that classifies putative TGEs by comparing to reference proteome sequences. This allows large-scale identification of sequence polymorphisms, splice isoforms and novel TGEs supported by presence or absence of variant-specific peptide evidence. Unlike previously reported methods, ours does not require a catalogue of known variants, making it more applicable to non-model organisms. The method was validated on human PIT data, then applied to Mus musculus, Pteropus alecto and Aedes aegypti. Novel discoveries included 60 human protein isoforms, 32 392 polymorphisms in P. alecto, and TGEs with non-methionine start sites including tyrosine.
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