Creaser Colin S, Green Philip S, Kilby Peter M, Ratcliffe Lucy
School of Biomedical and Natural Sciences, Nottingham Trent University, Nottingham NG11 8NS, UK.
Rapid Commun Mass Spectrom. 2006;20(5):829-36. doi: 10.1002/rcm.2376.
The atmospheric-pressure matrix-assisted laser desorption/ionisation quadrupole ion trap (AP-MALDI-QIT) analysis of tryptic peptides is reported following capillary liquid chromatographic (LC) separation and direct analysis of a protein digest. Peptide fragments were identified by peptide mass fingerprinting from mass spectrometric data and sequence analysis obtained by tandem mass spectrometry of the principal mass spectral peaks using a data-dependent scanning protocol. These data were compared with those from mass spectrometric analysis using capillary LC/MALDI-time-of-flight (TOF) and capillary LC/electrospray ionisation (ESI)-quadrupole TOF. For all three configurations the resulting data were searched against the MSDB database, using MASCOT and the sequence coverage compared for each technique. Complementary data were obtained using the three techniques.
本文报道了在毛细管液相色谱(LC)分离并直接分析蛋白质消化产物后,采用大气压基质辅助激光解吸/电离四极杆离子阱(AP-MALDI-QIT)对胰蛋白酶肽段进行分析的方法。通过肽质量指纹图谱从质谱数据中鉴定肽片段,并使用数据依赖扫描协议对主要质谱峰进行串联质谱分析以获得序列分析结果。将这些数据与使用毛细管LC/MALDI-飞行时间(TOF)和毛细管LC/电喷雾电离(ESI)-四极杆TOF进行质谱分析得到的数据进行比较。对于所有三种配置,使用MASCOT在MSDB数据库中搜索所得数据,并比较每种技术的序列覆盖率。使用这三种技术获得了互补数据。