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Conditions for Analysis of Native Protein Structures Using Uniform Field Drift Tube Ion Mobility Mass Spectrometry and Characterization of Stable Calibrants for TWIM-MS.
- Source :
-
Journal of the American Society for Mass Spectrometry [J Am Soc Mass Spectrom] 2019 Feb; Vol. 30 (2), pp. 256-267. Date of Electronic Publication: 2018 Oct 15. - Publication Year :
- 2019
-
Abstract
- Determination of collisional cross sections (CCS) by travelling wave ion mobility mass spectrometry (TWIM-MS) requires calibration against standards for which the CCS has been measured previously by drift tube ion mobility mass spectrometry (DTIM-MS). The different extents of collisional activation in TWIM-MS and DTIM-MS can give rise to discrepancies in the CCS of calibrants across the two platforms. Furthermore, the conditions required to ionize and transmit large, folded proteins and assemblies may variably affect the structure of the calibrants and analytes. Stable hetero-oligomeric phospholipase A <subscript>2</subscript> (PDx) and its subunits were characterized as calibrants for TWIM-MS. Conditions for acquisition of native-like TWIM (Synapt G1 HDMS) and DTIM (Agilent 6560 IM-Q-TOF) mass spectra were optimized to ensure the spectra exhibited similar charge state distributions. CCS measurements (DTIM-MS) for ubiquitin, cytochrome c, holo-myoglobin, serum albumin and glutamate dehydrogenase were in good agreement with other recent results determined using this and other DTIM-MS instruments. PDx and its β and γ subunits were stable across a wide range of cone and trap voltages in TWIM-MS and were stable in the presence of organic solvents. The CCS of PDx and its subunits were determined by DTIM-MS and were used as calibrants in determination of CCS of native-like cytochrome c, holo-myoglobin, carbonic anhydrase, serum albumin and haemoglobin in TWIM-MS. The CCS values were in good agreement with those measured by DTIM-MS where available. These experiments demonstrate conditions for analysis of native-like proteins using a commercially available DTIM-MS instrument, characterize robust calibrants for TWIM-MS, and present CCS values determined by DTIM-MS and TWIM-MS for native proteins to add to the current literature database. Graphical Abstract ᅟ.
- Subjects :
- Calibration
Carbonic Anhydrases analysis
Carbonic Anhydrases chemistry
Cytochromes c analysis
Cytochromes c chemistry
Myoglobin analysis
Myoglobin chemistry
Phospholipases A2 analysis
Phospholipases A2 chemistry
Protein Subunits
Proteins analysis
Serum Albumin, Human analysis
Serum Albumin, Human chemistry
Solvents chemistry
Spectrometry, Mass, Electrospray Ionization instrumentation
Spectrometry, Mass, Electrospray Ionization standards
Ubiquitin analysis
Ubiquitin chemistry
Ion Mobility Spectrometry methods
Proteins chemistry
Spectrometry, Mass, Electrospray Ionization methods
Subjects
Details
- Language :
- English
- ISSN :
- 1879-1123
- Volume :
- 30
- Issue :
- 2
- Database :
- MEDLINE
- Journal :
- Journal of the American Society for Mass Spectrometry
- Publication Type :
- Academic Journal
- Accession number :
- 30324262
- Full Text :
- https://doi.org/10.1007/s13361-018-2074-z