TY - JOUR
T1 - Signature ions in MS/MS spectra for dansyl-aminohexyl-QQIV adducts on lysine
AU - Schopfer, Lawrence M.
AU - Lockridge, Oksana
N1 - Funding Information:
Acknowledgments: Mass spectrometry data were obtained by the Mass Spectrometry and Proteomics Core Facility at the University of Nebraska Medical Center. We thank Robert J. Chalkley University of California San Francisco for helpful advice on the use of Protein Prospector software programs. Protein Prospector programs are available at no cost. http://prospector.ucsf.edu/prospector/mshome.htm. Programs were developed in the University of California San Francisco Mass Spectrometry Facility, directed by Alma Burlingame, funded by the NIH National Institute for General Medical Sciences. The Proteomics Toolkit http://db.systemsbiology.net:8080/proteomicsToolkit/ was used to identify ions in MS/MS spectra.
Funding Information:
Funding: Supported by the Fred and Pamela Buffet Cancer Center Support Grant P30CA036727 and NIH grant 1R21ES030132-01A1 (to O.L.).
Publisher Copyright:
© 2020 by the authors. Licensee MDPI, Basel, Switzerland.
PY - 2020/6
Y1 - 2020/6
N2 - Bacterial transglutaminase was used to label human plasma proteins with fluorescent tags. Protein lysines were modified with dansyl-epsilon-aminohexyl-Gln-Gln-Ile-Val-OH (dansylQQIV), while protein glutamines were modified with dansyl cadaverine. Labeled proteins included human butyrylcholinesterase, apolipoprotein A-1, haptoglobin, haptoglobin-related protein, immunoglobulin heavy chain, and hemopexin. Tryptic peptides were analyzed by LC-MS/MS on an Orbitrap Fusion Lumos mass spectrometer. Modified residues were identified in Protein Prospector and Proteome Discoverer searches of mass spectrometry data. The MS/MS fragmentation spectra from dansylQQIV-modified peptides gave intense peaks at 475.2015, 364.1691, 347.1426, 234.0585, and 170.0965 m/z. These signature ions are useful markers for identifying modified peptides. Human butyrylcholinesterase retained full activity following modification by dansylQQIV or dansyl cadaverine.
AB - Bacterial transglutaminase was used to label human plasma proteins with fluorescent tags. Protein lysines were modified with dansyl-epsilon-aminohexyl-Gln-Gln-Ile-Val-OH (dansylQQIV), while protein glutamines were modified with dansyl cadaverine. Labeled proteins included human butyrylcholinesterase, apolipoprotein A-1, haptoglobin, haptoglobin-related protein, immunoglobulin heavy chain, and hemopexin. Tryptic peptides were analyzed by LC-MS/MS on an Orbitrap Fusion Lumos mass spectrometer. Modified residues were identified in Protein Prospector and Proteome Discoverer searches of mass spectrometry data. The MS/MS fragmentation spectra from dansylQQIV-modified peptides gave intense peaks at 475.2015, 364.1691, 347.1426, 234.0585, and 170.0965 m/z. These signature ions are useful markers for identifying modified peptides. Human butyrylcholinesterase retained full activity following modification by dansylQQIV or dansyl cadaverine.
KW - Butyrylcholinesterase
KW - DansylQQIV
KW - Mass spectrometry
KW - Plasma proteins
KW - Protein Prospector
KW - Transglutaminase
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U2 - 10.3390/molecules25112659
DO - 10.3390/molecules25112659
M3 - Article
C2 - 32521655
AN - SCOPUS:85086355399
SN - 1420-3049
VL - 25
JO - Molecules
JF - Molecules
IS - 11
M1 - 2659
ER -