Detection of phosphatidylserine with a modified polar head group in human keratinocytes exposed to the radical generator AAPH

Elisabete Maciel, Bruno M. Neves, Deolinda Santinha, Ana Reis, Pedro Domingues, M. Teresa Cruz, Andrew R. Pitt, Corinne M. Spickett, M. Rosário M. Domingues

Research output: Contribution to journalArticle

Abstract

Phosphatidylserine (PS) is preferentially located in the inner leaflet of the cell membrane, and translocation of PS oxidized in fatty acyl chains to the outside of membrane has been reported as signaling to macrophage receptors to clear apoptotic cells. It was recently shown that PS can be oxidized in serine moiety of polar head-group. In the present work, a targeted lipidomic approach was applied to detecting OxPS modified at the polar head-group in keratinocytes that were exposed to the radical generator AAPH. Glycerophosphoacetic acid derivatives (GPAA) were found to be the major oxidation products of OxPS modified at the polar head-group during oxidation induced by AAPH-generated radicals, similarly to previous observations for the oxidation induced by OH radical. The neutral loss scan of 58Da and a novel precursor ion scan of m/z 137.1 (HOPO3CH2COOH) allowed the recognition of GPAA derivatives in the total lipid extracts obtained from HaCaT cells treated with AAPH. The positive identification of serine head group oxidation products in cells under controlled oxidative conditions opens new perspectives and justifies further studies in other cellular environments in order to understand fully the role of PS polar head-group oxidation in cell homeostasis and disease.
LanguageEnglish
Pages38–45
Number of pages8
JournalArchives of Biochemistry and Biophysics
Volume548
Early online date20 Feb 2014
DOIs
Publication statusPublished - 15 Apr 2014

Fingerprint

Phosphatidylserines
Keratinocytes
Oxidation
Serine
Derivatives
Acids
Macrophages
Cell membranes
Homeostasis
Cell Membrane
Ions
Lipids
Membranes
Cells
2,2'-azobis(2-amidinopropane)

Keywords

  • phosphatidylserine
  • keratinocytes
  • AAPH
  • oxidative stress
  • mass spectrometry
  • shotgun lipidomic

Cite this

Maciel, Elisabete ; Neves, Bruno M. ; Santinha, Deolinda ; Reis, Ana ; Domingues, Pedro ; Cruz, M. Teresa ; Pitt, Andrew R. ; Spickett, Corinne M. ; Domingues, M. Rosário M. / Detection of phosphatidylserine with a modified polar head group in human keratinocytes exposed to the radical generator AAPH. In: Archives of Biochemistry and Biophysics. 2014 ; Vol. 548. pp. 38–45.
@article{008727ae6b1d48fcadd0fd9d68d74479,
title = "Detection of phosphatidylserine with a modified polar head group in human keratinocytes exposed to the radical generator AAPH",
abstract = "Phosphatidylserine (PS) is preferentially located in the inner leaflet of the cell membrane, and translocation of PS oxidized in fatty acyl chains to the outside of membrane has been reported as signaling to macrophage receptors to clear apoptotic cells. It was recently shown that PS can be oxidized in serine moiety of polar head-group. In the present work, a targeted lipidomic approach was applied to detecting OxPS modified at the polar head-group in keratinocytes that were exposed to the radical generator AAPH. Glycerophosphoacetic acid derivatives (GPAA) were found to be the major oxidation products of OxPS modified at the polar head-group during oxidation induced by AAPH-generated radicals, similarly to previous observations for the oxidation induced by OH radical. The neutral loss scan of 58Da and a novel precursor ion scan of m/z 137.1 (HOPO3CH2COOH) allowed the recognition of GPAA derivatives in the total lipid extracts obtained from HaCaT cells treated with AAPH. The positive identification of serine head group oxidation products in cells under controlled oxidative conditions opens new perspectives and justifies further studies in other cellular environments in order to understand fully the role of PS polar head-group oxidation in cell homeostasis and disease.",
keywords = "phosphatidylserine, keratinocytes, AAPH, oxidative stress, mass spectrometry, shotgun lipidomic",
author = "Elisabete Maciel and Neves, {Bruno M.} and Deolinda Santinha and Ana Reis and Pedro Domingues and Cruz, {M. Teresa} and Pitt, {Andrew R.} and Spickett, {Corinne M.} and Domingues, {M. Ros{\'a}rio M.}",
year = "2014",
month = "4",
day = "15",
doi = "10.1016/j.abb.2014.02.002",
language = "English",
volume = "548",
pages = "38–45",
journal = "Archives of Biochemistry and Biophysics",
issn = "0003-9861",
publisher = "Academic Press Inc.",

}

Detection of phosphatidylserine with a modified polar head group in human keratinocytes exposed to the radical generator AAPH. / Maciel, Elisabete; Neves, Bruno M.; Santinha, Deolinda; Reis, Ana; Domingues, Pedro; Cruz, M. Teresa; Pitt, Andrew R.; Spickett, Corinne M.; Domingues, M. Rosário M.

In: Archives of Biochemistry and Biophysics, Vol. 548, 15.04.2014, p. 38–45.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Detection of phosphatidylserine with a modified polar head group in human keratinocytes exposed to the radical generator AAPH

AU - Maciel, Elisabete

AU - Neves, Bruno M.

AU - Santinha, Deolinda

AU - Reis, Ana

AU - Domingues, Pedro

AU - Cruz, M. Teresa

AU - Pitt, Andrew R.

AU - Spickett, Corinne M.

AU - Domingues, M. Rosário M.

PY - 2014/4/15

Y1 - 2014/4/15

N2 - Phosphatidylserine (PS) is preferentially located in the inner leaflet of the cell membrane, and translocation of PS oxidized in fatty acyl chains to the outside of membrane has been reported as signaling to macrophage receptors to clear apoptotic cells. It was recently shown that PS can be oxidized in serine moiety of polar head-group. In the present work, a targeted lipidomic approach was applied to detecting OxPS modified at the polar head-group in keratinocytes that were exposed to the radical generator AAPH. Glycerophosphoacetic acid derivatives (GPAA) were found to be the major oxidation products of OxPS modified at the polar head-group during oxidation induced by AAPH-generated radicals, similarly to previous observations for the oxidation induced by OH radical. The neutral loss scan of 58Da and a novel precursor ion scan of m/z 137.1 (HOPO3CH2COOH) allowed the recognition of GPAA derivatives in the total lipid extracts obtained from HaCaT cells treated with AAPH. The positive identification of serine head group oxidation products in cells under controlled oxidative conditions opens new perspectives and justifies further studies in other cellular environments in order to understand fully the role of PS polar head-group oxidation in cell homeostasis and disease.

AB - Phosphatidylserine (PS) is preferentially located in the inner leaflet of the cell membrane, and translocation of PS oxidized in fatty acyl chains to the outside of membrane has been reported as signaling to macrophage receptors to clear apoptotic cells. It was recently shown that PS can be oxidized in serine moiety of polar head-group. In the present work, a targeted lipidomic approach was applied to detecting OxPS modified at the polar head-group in keratinocytes that were exposed to the radical generator AAPH. Glycerophosphoacetic acid derivatives (GPAA) were found to be the major oxidation products of OxPS modified at the polar head-group during oxidation induced by AAPH-generated radicals, similarly to previous observations for the oxidation induced by OH radical. The neutral loss scan of 58Da and a novel precursor ion scan of m/z 137.1 (HOPO3CH2COOH) allowed the recognition of GPAA derivatives in the total lipid extracts obtained from HaCaT cells treated with AAPH. The positive identification of serine head group oxidation products in cells under controlled oxidative conditions opens new perspectives and justifies further studies in other cellular environments in order to understand fully the role of PS polar head-group oxidation in cell homeostasis and disease.

KW - phosphatidylserine

KW - keratinocytes

KW - AAPH

KW - oxidative stress

KW - mass spectrometry

KW - shotgun lipidomic

UR - http://www.scopus.com/inward/record.url?scp=84897009492&partnerID=8YFLogxK

U2 - 10.1016/j.abb.2014.02.002

DO - 10.1016/j.abb.2014.02.002

M3 - Article

VL - 548

SP - 38

EP - 45

JO - Archives of Biochemistry and Biophysics

T2 - Archives of Biochemistry and Biophysics

JF - Archives of Biochemistry and Biophysics

SN - 0003-9861

ER -