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Identification and Characterization of Human Proteoforms by Top-Down LC-21 Tesla FT-ICR Mass Spectrometry.

Title: Identification and Characterization of Human Proteoforms by Top-Down LC-21 Tesla FT-ICR Mass Spectrometry.
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Name(s): Anderson, Lissa C, author
DeHart, Caroline J, author
Kaiser, Nathan K, author
Fellers, Ryan T, author
Smith, Donald F, author
Greer, Joseph B, author
LeDuc, Richard D, author
Blakney, Greg T, author
Thomas, Paul M, author
Kelleher, Neil L, author
Hendrickson, Christopher L, author
Type of Resource: text
Genre: Journal Article
Text
Date Issued: 2017-02-03
Physical Form: computer
online resource
Extent: 1 online resource
Language(s): English
Abstract/Description: Successful high-throughput characterization of intact proteins from complex biological samples by mass spectrometry requires instrumentation capable of high mass resolving power, mass accuracy, sensitivity, and spectral acquisition rate. These limitations often necessitate the performance of hundreds of LC-MS/MS experiments to obtain reasonable coverage of the targeted proteome, which is still typically limited to molecular weights below 30 kDa. The National High Magnetic Field Laboratory (NHMFL) recently installed a 21 T FT-ICR mass spectrometer, which is part of the NHMFL FT-ICR User Facility and available to all qualified users. Here we demonstrate top-down LC-21 T FT-ICR MS/MS of intact proteins derived from human colorectal cancer cell lysate. We identified a combined total of 684 unique protein entries observed as 3238 unique proteoforms at a 1% false discovery rate, based on rapid, data-dependent acquisition of collision-induced and electron-transfer dissociation tandem mass spectra from just 40 LC-MS/MS experiments. Our identifications included 372 proteoforms with molecular weights over 30 kDa detected at isotopic resolution, which substantially extends the accessible mass range for high-throughput top-down LC-MS/MS.
Identifier: FSU_pmch_27936753 (IID), 10.1021/acs.jproteome.6b00696 (DOI), PMC5353847 (PMCID), 27936753 (RID), 27936753 (EID)
Keywords: 21 tesla, FT-ICR, FTMS, Top-down proteomics
Grant Number: P30 CA060553, P41 GM108569
Publication Note: This NIH-funded author manuscript originally appeared in PubMed Central at https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5353847.
Subject(s): Amino Acid Sequence
Colorectal Neoplasms/chemistry
Colorectal Neoplasms/pathology
Complex Mixtures/chemistry
Cyclotrons/instrumentation
Fourier Analysis
Humans
Mass Spectrometry/instrumentation
Mass Spectrometry/methods
Neoplasm Proteins/analysis
Proteome/analysis
Proteomics/instrumentation
Proteomics/methods
Persistent Link to This Record: http://purl.flvc.org/fsu/fd/FSU_pmch_27936753
Owner Institution: FSU
Is Part Of: Journal of proteome research.
1535-3907
Issue: iss. 2, vol. 16

Choose the citation style.
Anderson, L. C., DeHart, C. J., Kaiser, N. K., Fellers, R. T., Smith, D. F., Greer, J. B., … Hendrickson, C. L. (2017). Identification and Characterization of Human Proteoforms by Top-Down LC-21 Tesla FT-ICR Mass Spectrometry. Journal Of Proteome Research. Retrieved from http://purl.flvc.org/fsu/fd/FSU_pmch_27936753