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The Calcium-Modulated Structures of Calmodulin and S100b Proteins are Useful to Monitor Hydrogen/Deuterium Exchange Efficiency Using Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry
- Source :
- European Journal of Mass Spectrometry. 15:739-746
- Publication Year :
- 2009
- Publisher :
- SAGE Publications, 2009.
-
Abstract
- Hydrogen/deuterium exchange (HDX) using matrix-assisted laser desorption ionization time-of-flight (MALDI-ToF) mass spectrometry is a sensitive, salt-tolerant and high-throughput method useful to probe protein conformation and molecular interactions. However, a drawback of the MALDI HDX technique is that sample preparation methods can typically result in higher levels of artificial deuterium in-exchange and/or hydrogen back-exchange just prior to or during mass analysis; this may impair data reproducibility and impede structural and kinetic data interpretation. While methods to minimize effects of back-exchange during protein analyte deposition on MALDI plates have been reported, this study presents a readily available, highly sensitive protein control set to facilitate rapid MALDI HDX protocol workup. The Ca2+-induced solvent accessible surface area (ASA) changes of calmodulin (CaM) and S100 proteins were employed to monitor and optimize HDX protocol efficiency. Under non-stringent room temperature conditions, the Ca2+-induced deuterium exchange of CaM, ΔDca2+, MH+ shifts −17 Da to −24 Da, while S100 ΔDca2+ MH+ shifts +8 Da to +12 Da. By comparing the divergent CaM and S100 Ca2+-induced deuterium mass shift differences, HDX sample workup and MALDI plate spotting conditions can easily be monitored.
- Subjects :
- MALDI imaging
Chemistry
S100 Proteins
Analytical chemistry
Deuterium Exchange Measurement
S100 Calcium Binding Protein beta Subunit
General Medicine
Mass spectrometry
Recombinant Proteins
Atomic and Molecular Physics, and Optics
Sample preparation in mass spectrometry
Protein Structure, Tertiary
Surface-enhanced laser desorption/ionization
Matrix-assisted laser desorption/ionization
Calmodulin
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Humans
Hydrogen–deuterium exchange
Nerve Growth Factors
Time-of-flight mass spectrometry
Protein Structure, Quaternary
Spectroscopy
Subjects
Details
- ISSN :
- 17516838 and 14690667
- Volume :
- 15
- Database :
- OpenAIRE
- Journal :
- European Journal of Mass Spectrometry
- Accession number :
- edsair.doi.dedup.....a90f5f39159d750e8bda38c39f0e8717