A Targeted, Differential Top-Down Proteomic Methodology for Comparison of ApoA-I Proteoforms in Individuals with High and Low HDL Efflux Capacity

Henrique Dos Santos Seckler, Luca Fornelli, R Kannan Mutharasan, Colby S Thaxton, Ryan Fellers, Martha Daviglus, Allan Sniderman, Daniel Rader, Neil L Kelleher, Donald M Lloyd-Jones, Philip D Compton, John T Wilkins*

*Corresponding author for this work

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Top-down proteomics (TDP) allows precise determination/characterization of the different proteoforms derived from the expression of a single gene. In this study, we targeted apolipoprotein A-I (ApoA-I), a mediator of high-density-lipoprotein cholesterol efflux (HDL-E), which is inversely associated with coronary heart disease risk. Absolute ApoA-I concentration and allelic variation only partially explain interindividual HDL-E variation. Therefore, we hypothesize that differences in HDL-E are associated with the abundances of different ApoA-I proteoforms. Here, we present a targeted TDP methodology to characterize ApoA-I proteoforms in serum samples and compare their abundances between individuals. We characterized 18 ApoA-I proteoforms using selected-ion monitoring coupled to electron-transfer dissociation mass spectrometry. We then compared the abundances of these proteoforms between two groups of four participants, representing the individuals with highest and lowest HDL-E values within the Chicago Healthy Aging Study (n = 420). Six proteoforms showed significantly (p < 0.0005) higher intensity in high HDL-E individuals: canonical ApoA-I [fold difference (fd) = 1.17], carboxymethylated ApoA-I (fd = 1.24) and, with highest difference, four fatty acylated forms: palmitoylated (fd = 2.16), oleoylated (fd = 2.08), arachidonoylated (fd = 2.31) and one bearing two modifications: palmitoylation and truncation (fd = 2.13). These results demonstrate translational potential for targeted TDP in revealing, with high sensitivity, associations between interindividual proteoform variation and physiological differences underlying disease risk.

Original languageEnglish (US)
Pages (from-to)2156-2164
Number of pages9
JournalJournal of Proteome Research
Volume17
Issue number6
DOIs
StatePublished - Jun 1 2018

Fingerprint

Apolipoprotein A-I
Proteomics
HDL Cholesterol
Bearings (structural)
Lipoylation
LDL Cholesterol
Coronary Disease
Mass spectrometry
Mass Spectrometry
Genes
Aging of materials
Electrons
Ions
Monitoring
Serum

Keywords

  • ApoA-I
  • HDL efflux
  • acylations
  • apolipoproteins
  • atherosclerosis
  • cholesterol
  • palmitoylation
  • proteoforms
  • top-down proteomics

ASJC Scopus subject areas

  • Biochemistry
  • Chemistry(all)

Cite this

@article{8ec91d063b9843ac92194458153aca2a,
title = "A Targeted, Differential Top-Down Proteomic Methodology for Comparison of ApoA-I Proteoforms in Individuals with High and Low HDL Efflux Capacity",
abstract = "Top-down proteomics (TDP) allows precise determination/characterization of the different proteoforms derived from the expression of a single gene. In this study, we targeted apolipoprotein A-I (ApoA-I), a mediator of high-density-lipoprotein cholesterol efflux (HDL-E), which is inversely associated with coronary heart disease risk. Absolute ApoA-I concentration and allelic variation only partially explain interindividual HDL-E variation. Therefore, we hypothesize that differences in HDL-E are associated with the abundances of different ApoA-I proteoforms. Here, we present a targeted TDP methodology to characterize ApoA-I proteoforms in serum samples and compare their abundances between individuals. We characterized 18 ApoA-I proteoforms using selected-ion monitoring coupled to electron-transfer dissociation mass spectrometry. We then compared the abundances of these proteoforms between two groups of four participants, representing the individuals with highest and lowest HDL-E values within the Chicago Healthy Aging Study (n = 420). Six proteoforms showed significantly (p < 0.0005) higher intensity in high HDL-E individuals: canonical ApoA-I [fold difference (fd) = 1.17], carboxymethylated ApoA-I (fd = 1.24) and, with highest difference, four fatty acylated forms: palmitoylated (fd = 2.16), oleoylated (fd = 2.08), arachidonoylated (fd = 2.31) and one bearing two modifications: palmitoylation and truncation (fd = 2.13). These results demonstrate translational potential for targeted TDP in revealing, with high sensitivity, associations between interindividual proteoform variation and physiological differences underlying disease risk.",
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A Targeted, Differential Top-Down Proteomic Methodology for Comparison of ApoA-I Proteoforms in Individuals with High and Low HDL Efflux Capacity. / Seckler, Henrique Dos Santos; Fornelli, Luca; Mutharasan, R Kannan; Thaxton, Colby S; Fellers, Ryan; Daviglus, Martha; Sniderman, Allan; Rader, Daniel; Kelleher, Neil L; Lloyd-Jones, Donald M; Compton, Philip D; Wilkins, John T.

In: Journal of Proteome Research, Vol. 17, No. 6, 01.06.2018, p. 2156-2164.

Research output: Contribution to journalArticle

TY - JOUR

T1 - A Targeted, Differential Top-Down Proteomic Methodology for Comparison of ApoA-I Proteoforms in Individuals with High and Low HDL Efflux Capacity

AU - Seckler, Henrique Dos Santos

AU - Fornelli, Luca

AU - Mutharasan, R Kannan

AU - Thaxton, Colby S

AU - Fellers, Ryan

AU - Daviglus, Martha

AU - Sniderman, Allan

AU - Rader, Daniel

AU - Kelleher, Neil L

AU - Lloyd-Jones, Donald M

AU - Compton, Philip D

AU - Wilkins, John T

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AB - Top-down proteomics (TDP) allows precise determination/characterization of the different proteoforms derived from the expression of a single gene. In this study, we targeted apolipoprotein A-I (ApoA-I), a mediator of high-density-lipoprotein cholesterol efflux (HDL-E), which is inversely associated with coronary heart disease risk. Absolute ApoA-I concentration and allelic variation only partially explain interindividual HDL-E variation. Therefore, we hypothesize that differences in HDL-E are associated with the abundances of different ApoA-I proteoforms. Here, we present a targeted TDP methodology to characterize ApoA-I proteoforms in serum samples and compare their abundances between individuals. We characterized 18 ApoA-I proteoforms using selected-ion monitoring coupled to electron-transfer dissociation mass spectrometry. We then compared the abundances of these proteoforms between two groups of four participants, representing the individuals with highest and lowest HDL-E values within the Chicago Healthy Aging Study (n = 420). Six proteoforms showed significantly (p < 0.0005) higher intensity in high HDL-E individuals: canonical ApoA-I [fold difference (fd) = 1.17], carboxymethylated ApoA-I (fd = 1.24) and, with highest difference, four fatty acylated forms: palmitoylated (fd = 2.16), oleoylated (fd = 2.08), arachidonoylated (fd = 2.31) and one bearing two modifications: palmitoylation and truncation (fd = 2.13). These results demonstrate translational potential for targeted TDP in revealing, with high sensitivity, associations between interindividual proteoform variation and physiological differences underlying disease risk.

KW - ApoA-I

KW - HDL efflux

KW - acylations

KW - apolipoproteins

KW - atherosclerosis

KW - cholesterol

KW - palmitoylation

KW - proteoforms

KW - top-down proteomics

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