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探索转录组图谱:超越16号染色体蛋白质组注释的一步。

Surfing transcriptomic landscapes. A step beyond the annotation of chromosome 16 proteome.

作者信息

Segura Víctor, Medina-Aunon Juan Alberto, Mora Maria I, Martínez-Bartolomé Salvador, Abian Joaquín, Aloria Kerman, Antúnez Oreto, Arizmendi Jesús M, Azkargorta Mikel, Barceló-Batllori Silvia, Beaskoetxea Jabier, Bech-Serra Joan J, Blanco Francisco, Monteiro Mariana B, Cáceres David, Canals Francesc, Carrascal Monserrat, Casal José Ignacio, Clemente Felipe, Colomé Nuria, Dasilva Noelia, Díaz Paula, Elortza Félix, Fernández-Puente Patricia, Fuentes Manuel, Gallardo Oscar, Gharbi Severine I, Gil Concha, González-Tejedo Carmen, Hernáez María Luisa, Lombardía Manuel, Lopez-Lucendo Maria, Marcilla Miguel, Mato José M, Mendes Marta, Oliveira Eliandre, Orera Irene, Pascual-Montano Alberto, Prieto Gorka, Ruiz-Romero Cristina, Sánchez del Pino Manuel M, Tabas-Madrid Daniel, Valero Maria L, Vialas Vital, Villanueva Joan, Albar Juan Pablo, Corrales Fernando J

机构信息

ProteoRed-ISCIII, Center for Applied Medical Research (CIMA), University of Navarra , Pío XII, 55; Ed. CIMA, 31008 Pamplona, Spain.

出版信息

J Proteome Res. 2014 Jan 3;13(1):158-72. doi: 10.1021/pr400721r. Epub 2013 Nov 5.

Abstract

The Spanish team of the Human Proteome Project (SpHPP) marked the annotation of Chr16 and data analysis as one of its priorities. Precise annotation of Chromosome 16 proteins according to C-HPP criteria is presented. Moreover, Human Body Map 2.0 RNA-Seq and Encyclopedia of DNA Elements (ENCODE) data sets were used to obtain further information relative to cell/tissue specific chromosome 16 coding gene expression patterns and to infer the presence of missing proteins. Twenty-four shotgun 2D-LC-MS/MS and gel/LC-MS/MS MIAPE compliant experiments, representing 41% coverage of chromosome 16 proteins, were performed. Furthermore, mapping of large-scale multicenter mass spectrometry data sets from CCD18, MCF7, Jurkat, and Ramos cell lines into RNA-Seq data allowed further insights relative to correlation of chromosome 16 transcripts and proteins. Detection and quantification of chromosome 16 proteins in biological matrices by SRM procedures are also primary goals of the SpHPP. Two strategies were undertaken: one focused on known proteins, taking advantage of MS data already available, and the second, aimed at the detection of the missing proteins, is based on the expression of recombinant proteins to gather MS information and optimize SRM methods that will be used in real biological samples. SRM methods for 49 known proteins and for recombinant forms of 24 missing proteins are reported in this study.

摘要

人类蛋白质组计划西班牙团队(SpHPP)将16号染色体的注释和数据分析列为其优先事项之一。本文根据C-HPP标准对16号染色体蛋白质进行了精确注释。此外,还利用了人类基因组图谱2.0 RNA测序和DNA元件百科全书(ENCODE)数据集,以获取有关细胞/组织特异性16号染色体编码基因表达模式的更多信息,并推断缺失蛋白质的存在。开展了24项符合MIAPE标准的鸟枪法二维液相色谱-串联质谱(2D-LC-MS/MS)和凝胶/液相色谱-串联质谱(gel/LC-MS/MS)实验,覆盖了16号染色体蛋白质的41%。此外,将来自CCD18、MCF7、Jurkat和Ramos细胞系的大规模多中心质谱数据集映射到RNA测序数据中,有助于进一步了解16号染色体转录本与蛋白质之间的相关性。通过SRM程序检测和定量生物基质中的16号染色体蛋白质也是SpHPP的主要目标。采取了两种策略:一种策略聚焦于已知蛋白质,利用已有的质谱数据;另一种策略旨在检测缺失蛋白质,基于重组蛋白质的表达来收集质谱信息并优化将用于实际生物样品的SRM方法。本研究报告了针对49种已知蛋白质和24种缺失蛋白质重组形式的SRM方法。

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