Pull-down Assay on Streptavidin Beads and Surface Plasmon
Resonance Chips for SWATH-MS-based Interactomics

J 2018

Pull-down Assay on Streptavidin Beads and Surface Plasmon Resonance Chips for SWATH-MS-based Interactomics

MARYÁŠ, Josef, Jakub FAKTOR, Lenka ČÁPKOVÁ, Petr MÜLLER, Petr SKLÁDAL et. al.

Základní údaje

Originální název

Pull-down Assay on Streptavidin Beads and Surface Plasmon Resonance Chips for SWATH-MS-based Interactomics

Autoři

MARYÁŠ, Josef (203 Česká republika, domácí), Jakub FAKTOR (703 Slovensko, domácí), Lenka ČÁPKOVÁ (203 Česká republika, domácí), Petr MÜLLER (203 Česká republika), Petr SKLÁDAL (203 Česká republika, domácí) a Pavel BOUCHAL (203 Česká republika, garant, domácí)

Vydání

CANCER GENOMICS & PROTEOMICS, ATHENS, INT INST ANTICANCER RESEARCH, 2018, 1109-6535

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10608 Biochemistry and molecular biology

Stát vydavatele

Řecko

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 3.147

Kód RIV

RIV/00216224:14310/18:00101543

Organizační jednotka

Přírodovědecká fakulta

DOI

http://dx.doi.org/10.21873/cgp.20098

UT WoS

000444004800005

Klíčová slova anglicky

LC-SWATH-MS/MS; pull-down assay; PDLIM2; protein-protein interactions; SPR

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 12. 3. 2021 17:35, doc. Mgr. Pavel Bouchal, Ph.D.

Anotace

V originále

Background/Aim: Pul-down assay is a popular in vitro method for identification of physical interactors of selected proteins. Here, for the first time, we compared three conventional variants of pull-down assay with the streptavidin-modified surface plasmon resonance (SPR) chips for the detection of PDZ and LIM domain protein 2 (PDLIM2) interaction partners. Materials and Methods: PDLIM2 protein-protein interactions were analysed by three variants of pull-down assay on streptavidin beads using LC-MS/MS in "Sequential Window Acquisition of all Theoretical fragment ion spectra (SWATH)" mode and compared with LC-SWATHMS/MS data from SPR chips. Results: The results showed that (i) the use of SPR chip led to comparable data compared to on-column streptavidin beads, (ii) gravity flow and microflow in wash and elution steps provided better results than centrifugation, and (iii) type and concentration of detergent did not significantly affect the interactome data of cancer-associated PDLIM2. Conclusion: Our study supports further application of SPR-based affinity purification with SWATH mass spectrometry for reproducible and controlled characterization of cancer-associated interactomes.

Návaznosti

GA17-05957S, projekt VaV

Název: Evaluace nových potenciálních cílů a inhibitorů pro blokování vývoje metastáz u luminálních A nádorů prsu

Investor: Grantová agentura ČR, Evaluace nových potenciálních cílů a inhibitorů pro blokování vývoje metastáz u luminálních A nádorů prsu

MUNI/A/1100/2017, interní kód MU

Název: Podpora biochemického výzkumu v roce 2018

Investor: Masarykova univerzita, Podpora biochemického výzkumu v roce 2018, DO R. 2020_Kategorie A - Specifický výzkum - Studentské výzkumné projekty

Citovat

MARYÁŠ, Josef, Jakub FAKTOR, Lenka ČÁPKOVÁ, Petr MÜLLER, Petr SKLÁDAL a Pavel BOUCHAL. Pull-down Assay on Streptavidin Beads and Surface Plasmon Resonance Chips for SWATH-MS-based Interactomics. CANCER GENOMICS & PROTEOMICS. ATHENS: INT INST ANTICANCER RESEARCH, 2018, roč. 15, č. 5, s. 395-404. ISSN 1109-6535. Dostupné z: https://dx.doi.org/10.21873/cgp.20098.

@article{1484743,
   author = {Maryáš, Josef and Faktor, Jakub and Čápková, Lenka and Müller, Petr and Skládal, Petr and Bouchal, Pavel},
   article_location = {ATHENS},
   article_number = {5},
   doi = {http://dx.doi.org/10.21873/cgp.20098},
   keywords = {LC-SWATH-MS/MS; pull-down assay; PDLIM2; protein-protein interactions; SPR},
   language = {eng},
   issn = {1109-6535},
   journal = {CANCER GENOMICS & PROTEOMICS},
   title = {Pull-down Assay on Streptavidin Beads and Surface Plasmon Resonance Chips for SWATH-MS-based Interactomics},
   url = {https://cgp.iiarjournals.org/content/15/5/395},
   volume = {15},
   year = {2018}
}

TY  - JOUR
ID  - 1484743
AU  - Maryáš, Josef - Faktor, Jakub - Čápková, Lenka - Müller, Petr - Skládal, Petr - Bouchal, Pavel
PY  - 2018
TI  - Pull-down Assay on Streptavidin Beads and Surface Plasmon Resonance Chips for SWATH-MS-based Interactomics
JF  - CANCER GENOMICS & PROTEOMICS
VL  - 15
IS  - 5
SP  - 395-404
EP  - 395-404
PB  - INT INST ANTICANCER RESEARCH
SN  - 11096535
KW  - LC-SWATH-MS/MS
KW  - pull-down assay
KW  - PDLIM2
KW  - protein-protein interactions
KW  - SPR
UR  - https://cgp.iiarjournals.org/content/15/5/395
N2  - Background/Aim: Pul-down assay is a popular in vitro method for identification of physical interactors of selected proteins. Here, for the first time, we compared three conventional variants of pull-down assay with the streptavidin-modified surface plasmon resonance (SPR) chips for the detection of PDZ and LIM domain protein 2 (PDLIM2) interaction partners. Materials and Methods: PDLIM2 protein-protein interactions were analysed by three variants of pull-down assay on streptavidin beads using LC-MS/MS in "Sequential Window Acquisition of all Theoretical fragment ion spectra (SWATH)" mode and compared with LC-SWATHMS/MS data from SPR chips. Results: The results showed that (i) the use of SPR chip led to comparable data compared to on-column streptavidin beads, (ii) gravity flow and microflow in wash and elution steps provided better results than centrifugation, and (iii) type and concentration of detergent did not significantly affect the interactome data of cancer-associated PDLIM2. Conclusion: Our study supports further application of SPR-based affinity purification with SWATH mass spectrometry for reproducible and controlled characterization of cancer-associated interactomes.
ER  -

MARYÁŠ, Josef, Jakub FAKTOR, Lenka ČÁPKOVÁ, Petr MÜLLER, Petr SKLÁDAL a Pavel BOUCHAL. Pull-down Assay on Streptavidin Beads and Surface Plasmon Resonance Chips for SWATH-MS-based Interactomics. \textit{CANCER GENOMICS \&{}amp; PROTEOMICS}. ATHENS: INT INST ANTICANCER RESEARCH, 2018, roč.~15, č.~5, s.~395-404. ISSN~1109-6535. Dostupné z: https://dx.doi.org/10.21873/cgp.20098.

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