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Direct Mass Spectrometry Analysis of Protein Complexes and Intact Proteins up to >70 kDa from Tissue

Reputable Mentor II
Reputable Mentor II
Rian L. Griffiths, Albert Konijnenberg, Rosa Viner and Helen J. Cooper
Anal. Chem.2019, 91,11, 6962-6966
Native liquid extraction surface analysis (LESA) mass spectrometry allows direct analysis of folded proteins and protein complexes from biological substrates, such as dried blood spots and thin tissue sections, by use of native-like extraction/ionization solvents. Previously, we have demonstrated native LESA mass spectrometry of folded proteins up to 16 kDa as well as the 64 kDa hemoglobin tetramer, from mouse tissues. With denaturing LESA solvents, the highest mass protein detected in tissue to date is ∼37 kDa. Here, we demonstrate native LESA mass spectrometry by use of a Q Exactive UHMR Hybrid Quadrupole-Orbitrap (QE-UHMR) mass spectrometer, pushing the upper mass limit of proteins detected in tissue to >70 kDa. Moreover, a protein trimer of 42 kDa was detected and its stoichiometry confirmed by higher energy collision dissociation (HCD). The benefits of inclusion of detergents in the LESA sampling solvent are also demonstrated.
School of Biosciences, University of Birmingham, Edgbaston, Birmingham B15 2TT, U.K. Thermo Fisher Scientific, Achtseweg Noord 5, 5651 GG Eindhoven, The Netherlands Thermo Fisher Scientific, 355 River Oaks Parkway, San Jose, California 95134, United States
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‎11-09-2021 01:53 AM
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