Breuker Kathrin, McLafferty Fred W
Institute of Organic Chemistry and Center for Molecular Biosciences Innsbruck, University of Innsbruck, Innrain 52a, 6020 Innsbruck, Austria.
Proc Natl Acad Sci U S A. 2008 Nov 25;105(47):18145-52. doi: 10.1073/pnas.0807005105.
Mass spectrometry (MS) has been revolutionized by electrospray ionization (ESI), which is sufficiently "gentle" to introduce nonvolatile biomolecules such as proteins and nucleic acids (RNA or DNA) into the gas phase without breaking covalent bonds. Although in some cases noncovalent bonding can be maintained sufficiently for ESI/MS characterization of the solution structure of large protein complexes and native enzyme/substrate binding, the new gaseous environment can ultimately cause dramatic structural alterations. The temporal (picoseconds to minutes) evolution of native protein structure during and after transfer into the gas phase, as proposed here based on a variety of studies, can involve side-chain collapse, unfolding, and refolding into new, non-native structures. Control of individual experimental factors allows optimization for specific research objectives.
质谱分析法（MS）因电喷雾电离（ESI）而发生了变革，电喷雾电离足够“温和”，能够在不破坏共价键的情况下将蛋白质和核酸（RNA或DNA）等非挥发性生物分子引入气相。尽管在某些情况下，非共价键能够得到足够维持，以便对大型蛋白质复合物的溶液结构和天然酶/底物结合进行电喷雾电离/质谱表征，但新的气态环境最终可能会导致显著的结构改变。根据各种研究推测，天然蛋白质结构在转移到气相过程中及之后的时间演变（从皮秒到分钟）可能涉及侧链塌缩、展开以及重新折叠成新的非天然结构。控制各个实验因素可针对特定研究目标进行优化。
