Host-centric proteomics of stool, part 2 of 2

Soluble extra-cellular proteins and peptides from human and mouse stool were fractionated on a C-4 solid phase extraction cartidge, trypsin digested, C-18 clean up and identified with MS/MS on a LTQ-Velos Orbitrap. Data processing: The readRAW program was used to generate peak lists from the original data. All protein sequences were reversed and concatenated to the original forward-oriented proteins. Spectra were assigned to peptides using a semi-specific enzyme specificity with the Mascot search algorithm (version 2.3.01) using static carbamidomethylation of cysteines, differential oxidized methionines, 50ppm precursor mass tolerance, 2 miscleavages and 0.8Da fragment ion tolerance. Searched against a mouse or human database. The current dataset contains the human data and PXD000240 contains the mouse data.

从人类与小鼠粪便中提取的可溶性胞外蛋白质与肽段，经C-4固相萃取柱（solid phase extraction cartridge）分级分离后，依次进行胰蛋白酶酶解、C-18脱盐处理，并通过LTQ-Velos Orbitrap质谱仪完成串联质谱（MS/MS）鉴定。
数据处理：使用readRAW程序从原始数据中生成峰列表；将所有蛋白质序列反向后与原始正向蛋白质序列拼接构建数据库。采用Mascot搜索算法（版本2.3.01），以半特异性酶切规则将谱图匹配至肽段，参数设置包括：半胱氨酸的固定羧酰胺甲基化修饰、甲硫氨酸的可变氧化修饰、前体离子质量容忍度50ppm、允许2个酶切漏切位点以及0.8Da的碎片离子质量容忍度。以人类或小鼠蛋白质组数据库作为检索库进行搜索。本数据集包含人类样本数据，小鼠样本数据收录于PXD000240数据集。