Human serum proteins preseparated by electrophoresis or chromatography followed by tandem mass spectrometry
Human serum proteins preseparated by electrophoresis or chromatography followed by tandem mass spectrometry
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DOI：
10.1021/pr034039p
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发表时间：
2004-05-01
影响因子：
4.4
通讯作者：
Jackowski, G
中科院分区：
文献类型：
--
作者：
Marshall, J;Jankowski, A;Jackowski, G
比较了电泳和色谱样品制备，并共同检测到人血清中存在约600种蛋白质产物。分析了通过离子电泳、色谱法或两者的组合预分离的来自粗血清的蛋白质。用胰蛋白酶或胰凝乳蛋白酶消化蛋白质。还通过反相色谱法收集天然存在的肽。通过串联质谱法鉴定所得肽。通过激光将肽从金属芯片解吸到四极杆飞行时间质谱仪中，或者通过金属针在电压下从反相色谱电喷雾电离到离子阱质谱仪中。所有与血清相关的常见蛋白质都被检测到，两台质谱仪对丰富的血清蛋白质的身份达成一致。在消化前预分离血清蛋白显著提高了从血液中检测非常见蛋白的能力。电泳和色谱为基础的实验被发现是互补的。许多新的细胞蛋白以前不与血清被记录。
Electrophoretic and chromatographic sample preparations were compared and together detected the presence of some 600 types of protein products in human serum. Proteins from crude serum preseparated by ionic electrophoresis, chromatography, or a combination of both were analyzed. Proteins were digested with trypsin or chymotrypsin. Naturally occurring peptides were also collected by reversed-phase chromatography. The resulting peptides were identified by tandem mass spectrometry. The peptides were either desorbed by a laser from a metal chip into a quadrupole-time-of-flight mass spectrometer or ionized as an electro-spray from reversed-phase chromatography via a metal needle under voltage into an ion-trap mass spectrometer. All of the commonly known proteins associated with serum were detected, and the two mass spectrometers agreed on the identity of abundant serum proteins. Preseparation of serum proteins prior to digestion markedly enhanced the capacity to detect un-common proteins from blood. Electrophoretic- and chromatography-based experiments were found to be complementary. Many novel cellular proteins not previously associated with serum were recorded.