摘要:
为了比较碰撞诱导解离(collision induced dissociation,CID)和高能碰撞解离(high energy collision dissociation,HCD)两种离子裂解模式在蛋白质组学分析方面的差异,分别应用这两种模式对牛血清白蛋白和细胞裂解物进行分析。通过比较所鉴定到的多肽和蛋白数目,发现在牛血清白蛋白中,HCD 碎裂方法所得的谱图匹配率和多肽 Mascot 得分均较高,表明其质谱图质量较好。在复杂样品细胞裂解物分析中,CID 碎裂方法鉴定到的谱图数、多肽数和蛋白数目均较多,表明该模式的灵敏度较高；HCD 碎裂方法所得的谱图匹配率和 Mascot 得分均较高,表明其质谱图质量较好。因此,这两种模式均可用于大规模蛋白质组学分析,CID 模式可提供更高的灵敏度,而 HCD 模式则可获得更高质量的质谱谱图。
Abstract:
To comprehensively compare the difference of collision induced dissociation (CID)and high energy collision dissociation (HCD),the tryptic digests of bovine serum albumin (BSA)and cell lysates were analyzed by LTQ-Velos Orbitrap.The HCD frag-mentation identified more peptides and this mode showed higher MS/MS success rate and higher Mascot score in the tryptic digests of BSA.The results demonstrated that the HCD fragmentation has higher MS/MS spectra quality than CID.But in the tryptic digests of cell lysates,the CID fragmentation identified more MS/MS spectra,peptides and proteins,which indicated that it produces higher sensitivity.The HCD fragmenta-tion shows higher MS/MS success rate and higher Mascot score distribution,demon-strating higher quality of MS/MS spectrum.Therefore,based on these findings,both CID and HCD modes could be used for large-scale proteomic analysis.The CID mode has higher sensitivity,and HCD mode can achieve higher quality of MS/MS spectrum.
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