Comprehensive metalloprotein analysis from a single separation
record

a 2013

Comprehensive metalloprotein analysis from a single separation record

TOMALOVÁ, Iva; Pavla FOLTYNOVÁ; Viktor KANICKÝ a Jan PREISLER

Základní údaje

Originální název

Comprehensive metalloprotein analysis from a single separation record

Autoři

TOMALOVÁ, Iva; Pavla FOLTYNOVÁ; Viktor KANICKÝ a Jan PREISLER

Vydání

20th International Symposium on Electro- and Liquid Phase-Separation Techniques, 2013

Další údaje

Jazyk

angličtina

Typ výsledku

Konferenční abstrakt

Obor

10406 Analytical chemistry

Stát vydavatele

Španělsko

Utajení

není předmětem státního či obchodního tajemství

Označené pro přenos do RIV

Ano

Kód RIV

RIV/00216224:14740/13:00066469

Organizační jednotka

Středoevropský technologický institut

Klíčová slova anglicky

metalloproteins; separation; mass spectrometry; hyphenated techniques

Změněno: 12. 12. 2013 14:25, Mgr. Iva Benešová, Ph.D.

Anotace

V originále

Metals play a crucial role in physiology and pathology of biological systems. It has been estimated that the metalloproteins encompass about one third of all proteins; they are involved in the regulation of protein expression, metal transportation, homeostasis or detoxification process. A novel method for comprehensive multidimensional analysis of metalloproteins is presented here. This approach is based on an off-line coupling of a single micro-column separation run to both substrate-assisted laser desorption (SALD) inductively coupled plasma (ICP) mass spectrometry (MS) and matrix-assisted laser desorption/ionization mass spectrometry (MALDI) MS. The effluent fractions are collected on a custom-designed Au-coated polyethylene terephthalate glycol (PETG) sample target that is compatible to both MS methods. The whole concept is demonstrated on successive analysis of rabbit-liver metallothionein (MT) isoform mixture: A single separation record analyzed consecutively by MALDI MS and SALD ICP MS provides information about both molecular mass of present proteins and metal distribution and quantity, respectively. To take full advantage of the off-line coupling potential, two different MALDI matrices in solutions of acidic and neutral pH were applied at the adjacent spots corresponding to MT peak. Thus additional information can be obtained: While under acidic condition the information about protein apoforms is revealed, the metal-protein complexes can be detected at neutral pH. The conditions of separation system as well as the performance of the designed sample target in comparison with commonly used MALDI targets or uncoated PETG sheets are investigated in detail. We believe the presented method can be a viable alternative to on-line coupling employing electrospray ionization and nebulizer ICP MS.

Návaznosti

ED1.1.00/02.0068, projekt VaV

Název: CEITEC - central european institute of technology

GAP206/12/0538, projekt VaV

Název: Nové strategie úpravy vzorků pro desorpční hmotnostní spektrometrii

Investor: Grantová agentura ČR, Nové strategie úpravy vzorků pro desorpční hmotnostní spektrometrii

Citovat

TOMALOVÁ, Iva; Pavla FOLTYNOVÁ; Viktor KANICKÝ a Jan PREISLER. Comprehensive metalloprotein analysis from a single separation record. In 20th International Symposium on Electro- and Liquid Phase-Separation Techniques. 2013.

@proceedings{1127503,
   author = {Tomalová, Iva and Foltynová, Pavla and Kanický, Viktor and Preisler, Jan},
   booktitle = {20th International Symposium on Electro- and Liquid Phase-Separation Techniques},
   keywords = {metalloproteins; separation; mass spectrometry; hyphenated techniques},
   language = {eng},
   title = {Comprehensive metalloprotein analysis from a single separation record},
   year = {2013}
}

TY  - CONF
ID  - 1127503
AU  - Tomalová, Iva - Foltynová, Pavla - Kanický, Viktor - Preisler, Jan
PY  - 2013
TI  - Comprehensive metalloprotein analysis from a single separation record
KW  - metalloproteins
KW  - separation
KW  - mass spectrometry
KW  - hyphenated techniques
N2  - Metals play a crucial role in physiology and pathology of biological systems. It has been estimated that the metalloproteins encompass about one third of all proteins; they are involved in the regulation of protein expression, metal transportation, homeostasis or detoxification process. A novel method for comprehensive multidimensional analysis of metalloproteins is presented here. This approach is based on an off-line coupling of a single micro-column separation run to both substrate-assisted laser desorption (SALD) inductively coupled plasma (ICP) mass spectrometry (MS) and matrix-assisted laser desorption/ionization mass spectrometry (MALDI) MS. The effluent fractions are collected on a custom-designed Au-coated polyethylene terephthalate glycol (PETG) sample target that is compatible to both MS methods. The whole concept is demonstrated on successive analysis of rabbit-liver metallothionein (MT) isoform mixture: A single separation record analyzed consecutively by MALDI MS and SALD ICP MS provides information about both molecular mass of present proteins and metal distribution and quantity, respectively. To take full advantage of the off-line coupling potential, two different MALDI matrices in solutions of acidic and neutral pH were applied at the adjacent spots corresponding to MT peak. Thus additional information can be obtained: While under acidic condition the information about protein apoforms is revealed, the metal-protein complexes can be detected at neutral pH. The conditions of separation system as well as the performance of the designed sample target in comparison with commonly used MALDI targets or uncoated PETG sheets are investigated in detail. We believe the presented method can be a viable alternative to on-line coupling employing electrospray ionization and nebulizer ICP MS.
ER  -

TOMALOVÁ, Iva; Pavla FOLTYNOVÁ; Viktor KANICKÝ a Jan PREISLER. Comprehensive metalloprotein analysis from a single separation record. In \textit{20th International Symposium on Electro- and Liquid Phase-Separation Techniques}. 2013.

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