Hunt D F, Yates J R, Shabanowitz J, Winston S, Hauer C R
Proc Natl Acad Sci U S A. 1986 Sep;83(17):6233-7. doi: 10.1073/pnas.83.17.6233.
Methodology for determining amino acid sequences of proteins by tandem mass spectrometry is described. The approach involves enzymatic and/or chemical degradation of the protein to a collection of peptides which are then fractionated by high-performance liquid chromatography. Each fraction, containing as many as 10-15 peptides, is then analyzed directly, without further purification, by a combination of liquid secondary-ion/collision-activated dissociation mass spectrometry on a multianalyzer instrument. Interpretation of collision-activated dissociation mass spectra is described, and results are presented from a study of soluble peptides produced by treatment of apolipoprotein B with cyanogen bromide and trypsin.
描述了通过串联质谱法测定蛋白质氨基酸序列的方法。该方法包括将蛋白质进行酶促和/或化学降解，得到一系列肽段，然后通过高效液相色谱法对这些肽段进行分离。每个含有多达10 - 15个肽段的馏分，随后在多分析仪仪器上通过液相二次离子/碰撞激活解离质谱联用直接进行分析，无需进一步纯化。文中描述了对碰撞激活解离质谱的解读，并给出了对用溴化氰和胰蛋白酶处理载脂蛋白B产生的可溶性肽段进行研究的结果。
