Snyder Dalton T, Lin Yu-Fu, Somogyi Arpad, Wysocki Vicki
Resource for Native MS Guided Structural Biology, The Ohio State University, Columbus OH, USA 43210.
Department of Chemistry and Biochemistry, The Ohio State University, Columbus OH, USA 43210.
Int J Mass Spectrom. 2021 Mar;461. doi: 10.1016/j.ijms.2020.116503. Epub 2021 Jan 5.
We describe instrumentation for conducting tandem surface-induced dissociation (tSID) of native protein complexes on an ultrahigh resolution Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometer. The two stages of SID are accomplished with split lenses replacing the entrance lenses of the quadrupole mass filter (stage 1, referred to herein as SID-Q) and the collision cell (stage 2, Q-SID). After SID-Q, the scattered projectile ions and subcomplexes formed in transit traverse the 20 mm pre-filter prior to the mass-selecting quadrupole, providing preliminary insights into the SID fragmentation kinetics of noncovalent protein complexes. The isolated SID fragments (subcomplexes) are then fragmented by SID in the collision cell entrance lens (Q-SID), generating subcomplexes of subcomplexes. We show that the ultrahigh resolution of the FT-ICR can be used for deconvolving species overlapping in which are particularly prominent in tandem SID spectra due to the combination of symmetric charge partitioning and narrow product ion charge state distributions. Various protein complex topologies are explored, including homotetramers, homopentamers, a homohexamer, and a heterohexamer.
我们描述了用于在超高分辨率傅里叶变换离子回旋共振（FT-ICR）质谱仪上对天然蛋白质复合物进行串联表面诱导解离（tSID）的仪器装置。SID的两个阶段通过替换四极质量过滤器入口透镜的分裂透镜来完成（第一阶段，在此称为SID-Q）和碰撞池（第二阶段，Q-SID）。在SID-Q之后，散射的入射离子和在传输过程中形成的亚复合物在质量选择四极之前穿过20毫米预过滤器，从而对非共价蛋白质复合物的SID碎片化动力学提供初步见解。然后，分离出的SID片段（亚复合物）在碰撞池入口透镜（Q-SID）中通过SID进行碎片化，生成亚复合物的亚复合物。我们表明，FT-ICR的超高分辨率可用于解卷积重叠的物种，这在串联SID光谱中尤为突出，这是由于对称电荷分配和窄产物离子电荷态分布的结合。我们探索了各种蛋白质复合物拓扑结构，包括同四聚体、同五聚体、同六聚体和异六聚体。
