A targeted proteomic assay for the measurement of plasma proteoforms related to human aging phenotypes

Richard David Semba, Pingbo Zhang, Min Zhu, Elisa Fabbri, Marta Gonzalez-Freire, Ruin Moaddel, Minghui Geng-Spyropoulos, Luigi Ferrucci

Research output: Contribution to journalArticle

Abstract

Circulating polypeptides and proteins have been implicated in reversing or accelerating aging phenotypes, including growth/differentiation factor 8 (GDF8), GDF11, eotaxin, and oxytocin. These proteoforms, which are defined as the protein products arising from a single gene due to alternative splicing and PTMs, have been challenging to study. Both GDF8 and GDF11 have known antagonists such as follistatin (FST), and WAP, Kazal, immunoglobulin, Kunitz, and NTR domain-containing proteins 1 and 2 (WFIKKN1, WFIKKN2). We developed a novel multiplexed SRM assay using LC-MS/MS to measure five proteins related to GDF8 and GDF11 signaling, and in addition, eotaxin, and oxytocin. Eighteen peptides consisting of 54 transitions were monitored and validated in pooled human plasma. In 24 adults, the mean (SD) concentrations (ng/mL) were as follows: GDF8 propeptide, 11.0 (2.4); GDF8 mature protein, 25.7 (8.0); GDF11 propeptide, 21.3 (10.9); GDF11 mature protein, 16.5 (12.4); FST, 29.8 (7.1); FST cleavage form FST303, 96.4 (69.2); WFIKKN1, 38.3 (8.3); WFIKKN2, 32.2 (10.5); oxytocin, 1.9 (0.9); and eotaxin, 2.3 (0.5). This novel multiplexed SRM assay should facilitate the study of the relationships of these proteoforms with major aging phenotypes.

Original languageEnglish (US)
Article number1600232
JournalProteomics
Volume17
Issue number15-16
DOIs
StatePublished - Aug 1 2017

Fingerprint

Myostatin
Proteomics
Follistatin
Assays
Aging of materials
Phenotype
Plasmas
Oxytocin
Proteins
Peptides
Pulse time modulation
Plasma (human)
Alternative Splicing
Immunoglobulins
Genes

Keywords

  • Aging
  • Eotaxin
  • Follistatin
  • Growth/differentiation factor 11
  • Growth/differentiation factor 8
  • Oxytocin
  • WFIKKN1
  • WFIKKN2

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

A targeted proteomic assay for the measurement of plasma proteoforms related to human aging phenotypes. / Semba, Richard David; Zhang, Pingbo; Zhu, Min; Fabbri, Elisa; Gonzalez-Freire, Marta; Moaddel, Ruin; Geng-Spyropoulos, Minghui; Ferrucci, Luigi.

In: Proteomics, Vol. 17, No. 15-16, 1600232, 01.08.2017.

Research output: Contribution to journalArticle

Semba, RD, Zhang, P, Zhu, M, Fabbri, E, Gonzalez-Freire, M, Moaddel, R, Geng-Spyropoulos, M & Ferrucci, L 2017, 'A targeted proteomic assay for the measurement of plasma proteoforms related to human aging phenotypes', Proteomics, vol. 17, no. 15-16, 1600232. https://doi.org/10.1002/pmic.201600232
Semba, Richard David ; Zhang, Pingbo ; Zhu, Min ; Fabbri, Elisa ; Gonzalez-Freire, Marta ; Moaddel, Ruin ; Geng-Spyropoulos, Minghui ; Ferrucci, Luigi. / A targeted proteomic assay for the measurement of plasma proteoforms related to human aging phenotypes. In: Proteomics. 2017 ; Vol. 17, No. 15-16.
@article{a6e31e361ee4461498d8e6a4044e7178,
title = "A targeted proteomic assay for the measurement of plasma proteoforms related to human aging phenotypes",
abstract = "Circulating polypeptides and proteins have been implicated in reversing or accelerating aging phenotypes, including growth/differentiation factor 8 (GDF8), GDF11, eotaxin, and oxytocin. These proteoforms, which are defined as the protein products arising from a single gene due to alternative splicing and PTMs, have been challenging to study. Both GDF8 and GDF11 have known antagonists such as follistatin (FST), and WAP, Kazal, immunoglobulin, Kunitz, and NTR domain-containing proteins 1 and 2 (WFIKKN1, WFIKKN2). We developed a novel multiplexed SRM assay using LC-MS/MS to measure five proteins related to GDF8 and GDF11 signaling, and in addition, eotaxin, and oxytocin. Eighteen peptides consisting of 54 transitions were monitored and validated in pooled human plasma. In 24 adults, the mean (SD) concentrations (ng/mL) were as follows: GDF8 propeptide, 11.0 (2.4); GDF8 mature protein, 25.7 (8.0); GDF11 propeptide, 21.3 (10.9); GDF11 mature protein, 16.5 (12.4); FST, 29.8 (7.1); FST cleavage form FST303, 96.4 (69.2); WFIKKN1, 38.3 (8.3); WFIKKN2, 32.2 (10.5); oxytocin, 1.9 (0.9); and eotaxin, 2.3 (0.5). This novel multiplexed SRM assay should facilitate the study of the relationships of these proteoforms with major aging phenotypes.",
keywords = "Aging, Eotaxin, Follistatin, Growth/differentiation factor 11, Growth/differentiation factor 8, Oxytocin, WFIKKN1, WFIKKN2",
author = "Semba, {Richard David} and Pingbo Zhang and Min Zhu and Elisa Fabbri and Marta Gonzalez-Freire and Ruin Moaddel and Minghui Geng-Spyropoulos and Luigi Ferrucci",
year = "2017",
month = "8",
day = "1",
doi = "10.1002/pmic.201600232",
language = "English (US)",
volume = "17",
journal = "Proteomics",
issn = "1615-9853",
publisher = "Wiley-VCH Verlag",
number = "15-16",

}

TY - JOUR

T1 - A targeted proteomic assay for the measurement of plasma proteoforms related to human aging phenotypes

AU - Semba, Richard David

AU - Zhang, Pingbo

AU - Zhu, Min

AU - Fabbri, Elisa

AU - Gonzalez-Freire, Marta

AU - Moaddel, Ruin

AU - Geng-Spyropoulos, Minghui

AU - Ferrucci, Luigi

PY - 2017/8/1

Y1 - 2017/8/1

N2 - Circulating polypeptides and proteins have been implicated in reversing or accelerating aging phenotypes, including growth/differentiation factor 8 (GDF8), GDF11, eotaxin, and oxytocin. These proteoforms, which are defined as the protein products arising from a single gene due to alternative splicing and PTMs, have been challenging to study. Both GDF8 and GDF11 have known antagonists such as follistatin (FST), and WAP, Kazal, immunoglobulin, Kunitz, and NTR domain-containing proteins 1 and 2 (WFIKKN1, WFIKKN2). We developed a novel multiplexed SRM assay using LC-MS/MS to measure five proteins related to GDF8 and GDF11 signaling, and in addition, eotaxin, and oxytocin. Eighteen peptides consisting of 54 transitions were monitored and validated in pooled human plasma. In 24 adults, the mean (SD) concentrations (ng/mL) were as follows: GDF8 propeptide, 11.0 (2.4); GDF8 mature protein, 25.7 (8.0); GDF11 propeptide, 21.3 (10.9); GDF11 mature protein, 16.5 (12.4); FST, 29.8 (7.1); FST cleavage form FST303, 96.4 (69.2); WFIKKN1, 38.3 (8.3); WFIKKN2, 32.2 (10.5); oxytocin, 1.9 (0.9); and eotaxin, 2.3 (0.5). This novel multiplexed SRM assay should facilitate the study of the relationships of these proteoforms with major aging phenotypes.

AB - Circulating polypeptides and proteins have been implicated in reversing or accelerating aging phenotypes, including growth/differentiation factor 8 (GDF8), GDF11, eotaxin, and oxytocin. These proteoforms, which are defined as the protein products arising from a single gene due to alternative splicing and PTMs, have been challenging to study. Both GDF8 and GDF11 have known antagonists such as follistatin (FST), and WAP, Kazal, immunoglobulin, Kunitz, and NTR domain-containing proteins 1 and 2 (WFIKKN1, WFIKKN2). We developed a novel multiplexed SRM assay using LC-MS/MS to measure five proteins related to GDF8 and GDF11 signaling, and in addition, eotaxin, and oxytocin. Eighteen peptides consisting of 54 transitions were monitored and validated in pooled human plasma. In 24 adults, the mean (SD) concentrations (ng/mL) were as follows: GDF8 propeptide, 11.0 (2.4); GDF8 mature protein, 25.7 (8.0); GDF11 propeptide, 21.3 (10.9); GDF11 mature protein, 16.5 (12.4); FST, 29.8 (7.1); FST cleavage form FST303, 96.4 (69.2); WFIKKN1, 38.3 (8.3); WFIKKN2, 32.2 (10.5); oxytocin, 1.9 (0.9); and eotaxin, 2.3 (0.5). This novel multiplexed SRM assay should facilitate the study of the relationships of these proteoforms with major aging phenotypes.

KW - Aging

KW - Eotaxin

KW - Follistatin

KW - Growth/differentiation factor 11

KW - Growth/differentiation factor 8

KW - Oxytocin

KW - WFIKKN1

KW - WFIKKN2

UR - http://www.scopus.com/inward/record.url?scp=85027890129&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85027890129&partnerID=8YFLogxK

U2 - 10.1002/pmic.201600232

DO - 10.1002/pmic.201600232

M3 - Article

VL - 17

JO - Proteomics

JF - Proteomics

SN - 1615-9853

IS - 15-16

M1 - 1600232

ER -