The dominance of arginine-containing peptides in MALDI-derived tryptic mass fingerprints of proteins
The dominance of arginine-containing peptides in MALDI-derived tryptic mass fingerprints of proteins
复制标题
DOI：
10.1021/ac990298f
复制
发表时间：
1999-10-01
影响因子：
7.4
通讯作者：
Jungblut, PR
中科院分区：
文献类型：
--
作者：
Krause, E;Wenschuh, H;Jungblut, PR
基质辅助激光解吸/电离质谱 (MALDI-MS) 是一种强大的工具，用于对通过二维电泳 (2-DE) 分离的蛋白质进行酶促胶内消化后获得的肽混合物进行质量指纹分析。在使用质谱鉴定对分枝杆菌进行蛋白质组分析的过程中，发现 94% 最强的 MALDI-MS 峰表示 C 末端带有精氨酸的肽。使用已知存在于分枝杆菌 35 kDa 抗原胰蛋白酶消化物中的合成肽的等摩尔混合物，证明该效果同样显着（“合成质量图”）。此外，还检查了仅在 C 末端（Arg 或 Lys）不同的合成肽的几种二元混合物，以合理化对含精氨酸肽的更高敏感性。所描述的影响程度取决于所使用的矩阵，并且可能有助于将质量指纹数据更可靠地分配给数据库中的蛋白质序列。
Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) is a powerful tool for mass fingerprinting of peptide mixtures obtained after enzymatic in-gel digestion of proteins separated by two-dimensional electrophoresis (2-DE). In the course of a proteome analysis of mycobacteria using mass spectrometric identification, it was found that 94% of the most intense MALDI-MS peaks denote peptides bearing arginine at the C-terminal end. The effect was demonstrated to be equally prominent using an equimolar mixture of the synthetic peptides known to be present in the tryptic digest of the mycobacterial 35 kDa antigen ("synthetic mass map"). In addition, several binary mixtures of synthetic peptides differing exclusively at the C terminus (Arg or Lys) were examined to rationalize the higher sensitivity toward arginine-containing peptides. The extent of the effect described depends on the matrix used and may facilitate a more reliable assignment of mass fingerprint data to protein sequences in databases.