10.1021/acs.jproteome.5b00270.s001
Anna Heink
Anna
Heink
W. Sean Davidson
W. Sean
Davidson
Debi K. Swertfeger
Debi K.
Swertfeger
L. Jason Lu
L. Jason
Lu
Amy S. Shah
Amy S.
Shah
A Comparison of
Methods To Enhance Protein Detection
of Lipoproteins by Mass Spectrometry
American Chemical Society
2015
Enhance Protein Detection
mass spectrometry
gel filtration chromatography
method
AALS
sequence coverage
mass spectrometry analysis
LRA
OSD
lipid removal agent
density lipoprotein size range
2015-07-02 00:00:00
Journal contribution
https://acs.figshare.com/articles/journal_contribution/A_Comparison_of_Methods_To_Enhance_Protein_Detection_of_Lipoproteins_by_Mass_Spectrometry/2153683
We sought to develop a new method
to more efficiently analyze lipid-bound
proteins by mass spectrometry using a combination of a lipid removal
agent (LRA) that selectively targets lipid-bound proteins and a mass
spectrometry compatible detergent, anionic acid labile surfactant
(AALS), that is capable of eluting proteins off the LRA. This method
was compared to established methods that use the lipid removal agent
alone and straight proteomic analysis of human plasma after organic
solvent delipidation (OSD). Plasma from healthy individuals was separated
by gel filtration chromatography and prepared for mass spectrometry
analysis by each of the described methods. The addition of AALS to
LRA increased the overall number of proteins detected in both the
high and low density lipoprotein size range, the number of peptide
counts for each protein, and the overall sequence coverage. Organic
solvent delipidation detected the most proteins, though with some
decrease in overall protein detection and sequence coverage due to
the presence of nonlipid-bound proteins. The use of LRA allows for
selection and analysis of lipid-bound proteins. The addition of a
mass spectrometry compatible detergent improved detection of lipid-bound
proteins from human plasma using LRA.