Identification of selenium-containing proteins in HEK 293 kidney cells using multiple chromatographies, LC-ICPMS and nano-LC-ESIMS

Karnakar R. Chitta; Julio A. Landero-Figueroa; Phanichand Kodali; Joseph A. Caruso; Edward J. Merino

doi:10.1016/j.talanta.2013.03.003

Identification of selenium-containing proteins in HEK 293 kidney cells using multiple chromatographies, LC-ICPMS and nano-LC-ESIMS

Karnakar R. Chitta, Julio A. Landero-Figueroa, Phanichand Kodali, Joseph A. Caruso, Edward J. Merino

Research output: Contribution to journal › Article › peer-review

9 Scopus citations

Abstract

Our previous studies using HeLa and HEK 293 cells demonstrated that selenomethionine, SeMet, exerts more of an antagonistic effect on arsenic than other selenium species. These studies attributed the antagonistic effect of SeMet to decreased levels of reactive oxygen species, ROS, changes in protein phosphorylation and possible incorporation of SeMet into proteins. The present study employs a metallomics approach to identify the selenium-containing proteins in HEK 293 cells raised with SeMet. The proteins were screened and separated using two dimensional high performance liquid chromatography (HPLC)-inductively coupled plasma mass spectrometry (ICPMS), size exclusion chromatography (SEC) and reversed-phase chromatography (RPC). The Se-containing proteins were identified by peptide mapping using nano-HPLC-Chip-electrospray ionization mass spectrometry (ESIMS).

Original language	English
Pages (from-to)	25-31
Number of pages	7
Journal	Talanta
Volume	114
DOIs	https://doi.org/10.1016/j.talanta.2013.03.003
State	Published - 2013
Externally published	Yes

Keywords

Identifying selenium-containing proteins
LC-ESIMS
Metallomics
Multiple chromatographic separations
SEC-ICPMS

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10.1016/j.talanta.2013.03.003

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title = "Identification of selenium-containing proteins in HEK 293 kidney cells using multiple chromatographies, LC-ICPMS and nano-LC-ESIMS",

abstract = "Our previous studies using HeLa and HEK 293 cells demonstrated that selenomethionine, SeMet, exerts more of an antagonistic effect on arsenic than other selenium species. These studies attributed the antagonistic effect of SeMet to decreased levels of reactive oxygen species, ROS, changes in protein phosphorylation and possible incorporation of SeMet into proteins. The present study employs a metallomics approach to identify the selenium-containing proteins in HEK 293 cells raised with SeMet. The proteins were screened and separated using two dimensional high performance liquid chromatography (HPLC)-inductively coupled plasma mass spectrometry (ICPMS), size exclusion chromatography (SEC) and reversed-phase chromatography (RPC). The Se-containing proteins were identified by peptide mapping using nano-HPLC-Chip-electrospray ionization mass spectrometry (ESIMS).",

keywords = "Identifying selenium-containing proteins, LC-ESIMS, Metallomics, Multiple chromatographic separations, SEC-ICPMS",

author = "Chitta, {Karnakar R.} and Landero-Figueroa, {Julio A.} and Phanichand Kodali and Caruso, {Joseph A.} and Merino, {Edward J.}",

year = "2013",

doi = "10.1016/j.talanta.2013.03.003",

language = "English",

volume = "114",

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journal = "Talanta",

issn = "0039-9140",

publisher = "Elsevier B.V.",

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T1 - Identification of selenium-containing proteins in HEK 293 kidney cells using multiple chromatographies, LC-ICPMS and nano-LC-ESIMS

AU - Chitta, Karnakar R.

AU - Landero-Figueroa, Julio A.

AU - Kodali, Phanichand

AU - Caruso, Joseph A.

AU - Merino, Edward J.

PY - 2013

Y1 - 2013

N2 - Our previous studies using HeLa and HEK 293 cells demonstrated that selenomethionine, SeMet, exerts more of an antagonistic effect on arsenic than other selenium species. These studies attributed the antagonistic effect of SeMet to decreased levels of reactive oxygen species, ROS, changes in protein phosphorylation and possible incorporation of SeMet into proteins. The present study employs a metallomics approach to identify the selenium-containing proteins in HEK 293 cells raised with SeMet. The proteins were screened and separated using two dimensional high performance liquid chromatography (HPLC)-inductively coupled plasma mass spectrometry (ICPMS), size exclusion chromatography (SEC) and reversed-phase chromatography (RPC). The Se-containing proteins were identified by peptide mapping using nano-HPLC-Chip-electrospray ionization mass spectrometry (ESIMS).

AB - Our previous studies using HeLa and HEK 293 cells demonstrated that selenomethionine, SeMet, exerts more of an antagonistic effect on arsenic than other selenium species. These studies attributed the antagonistic effect of SeMet to decreased levels of reactive oxygen species, ROS, changes in protein phosphorylation and possible incorporation of SeMet into proteins. The present study employs a metallomics approach to identify the selenium-containing proteins in HEK 293 cells raised with SeMet. The proteins were screened and separated using two dimensional high performance liquid chromatography (HPLC)-inductively coupled plasma mass spectrometry (ICPMS), size exclusion chromatography (SEC) and reversed-phase chromatography (RPC). The Se-containing proteins were identified by peptide mapping using nano-HPLC-Chip-electrospray ionization mass spectrometry (ESIMS).

KW - Identifying selenium-containing proteins

KW - LC-ESIMS

KW - Metallomics

KW - Multiple chromatographic separations

KW - SEC-ICPMS

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U2 - 10.1016/j.talanta.2013.03.003

DO - 10.1016/j.talanta.2013.03.003

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JO - Talanta

JF - Talanta

ER -

Identification of selenium-containing proteins in HEK 293 kidney cells using multiple chromatographies, LC-ICPMS and nano-LC-ESIMS

Abstract

Keywords

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