PhD Scientific Days 2023

Molecular Sciences - Posters B

Critical re-investigations of alkylation reactions in relation to the detection of per- and polysulfide species in cells

Előadó neve

Jurányi, Eszter Petra

Neptun code

ATLYXV

Előadó munkahelye

National Institute of Oncology

Előadó telefonszáma

06202696889

Előadó e-mail címe

juranyi.e.petra@gmail.com

Az előadás címe

Critical re-investigations of alkylation reactions in relation to the detection of per- and polysulfide species in cells

Szerző(k) neve és munkahelye

Eszter Petra Jurányi1,2, Klaudia Galambos1,3, Tamás Ditrói1, Péter Nagy1,4,5
1Department of Molecular Immunology and Toxicology and National Tumor Biology Laboratory, National Institute of Oncology, Budapest, Hungary
2Doctoral School of Molecular Medicine, Semmelweis University, Budapest, Hungary
3Laki Kálmán Doctoral School, University of Debrecen, Debrecen, Hungary
4Department of Anatomy and Histology, Laboratory of Redox Biology, University of Veterinary Medicine, Budapest, Hungary
5Institute of Chemistry, University of Debrecen, Debrecen, Hungary

Bemutatás módja

Poszter

Szekció

Molecular Sciences - Posters B

Language of the presentation

English

Preferred session

Molecular Sciences

Összefoglaló szövege

Introduction: The detection of Reactive Sulfur Species (RSS), especially low and high molecular weight per- and polysulfide species is a challenging task, because these molecules are quite reactive and labile species and present in relatively low concentrations inside living organisms. Their roles inside living cells however are very important, as they can protect against oxidative stress and regulate protein functions.
Aims: It is necessary to have a reliable, robust, accurate method for the quantitation of different persulfide species. While the numerous different published persulfide detection methods rely on different approaches, almost all of them employ some kind of alkylation step to conserve the persulfidome inside the sample at a given moment. In our work we have focused on this alkylation step and tried to carefully and systematically revise critical parameters affecting the reaction
Method: For this task we have chosen an alkylating agent especially recommended for persulfide detection, an iodoacetamide derivative β-hydroxy-4-phenylethyl-iodoacetamide (HPE-IAM) that has been shown to protect against polysulfide chain cleavage after alkylation. (Hamid et al. Red.Biol. 2019).
Results: We have re-investigated the alkyaltion based HPE-IAM method for the measurement of low molecular weight reactive sulfur species. We identified critical points of the protocol affecting the accuracy and reproducibility of detection. We have confirmed, that the yield of the alkylation reaction is sensitive to changes in temperature and alkylation time, but a tightly controlled sample preparation procedure allow the reaction to be carried out in a highly reproducible manner.
Conclusion: Our results highlight the importance of a well validated and carefully executed protocol and the possible problems associated with the alkylation reaction of persulfides.
Funding: The project was implemented with the support from the National Research, Development and Innovation Fund of the Ministry of Culture and Innovation under the National Laboratories Program (National Tumor Biology Laboratory (2022-2.1.1-NL-2022-00010)) and the Hungarian Thematic Excellence Program (under project TKP2021-EGA-44) Grant Agreements with the National Research, Development and Innovation Office.

University and Doctoral School

Semmelweis University, Doctoral School of Molecular Medicine

Supervisor

Prof. Péter Nagy

Publication of my abstract

I do not give consent to the publication of my abstract on the website of the congress.

Kind

Szabad

Status

elfogadva

Accepted presentation method

poszter

Előadás fájl jóváhagyás

nem rendelkezett róla

Előadó

7402

Start

11:54

End

11:59

Authors (legacy)

Eszter Petra Jurányi1,2, Klaudia Galambos1,3, Tamás Ditrói1, Péter Nagy1,4,5
1Department of Molecular Immunology and Toxicology and National Tumor Biology Laboratory, National Institute of Oncology, Budapest, Hungary
2Doctoral School of Molecular Medicine, Semmelweis University, Budapest, Hungary
3Laki Kálmán Doctoral School, University of Debrecen, Debrecen, Hungary
4Department of Anatomy and Histology, Laboratory of Redox Biology, University of Veterinary Medicine, Budapest, Hungary
5Institute of Chemistry, University of Debrecen, Debrecen, Hungary