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Pregled bibliografske jedinice broj: 767751

Plant seryl-tRNA synthetase and metabolic protein BEN1: identification of interaction surfaces by biophysical methods


Kekez, Mario; Zanki, Vladimir; Hodnik, Vesna; Anderluh, Gregor; Rokov Plavec, Jasmina
Plant seryl-tRNA synthetase and metabolic protein BEN1: identification of interaction surfaces by biophysical methods // Symposium & Workshop on Microscale Thermophoresis: Book of Abstracts / Godinić Mikulčić, Vlatka ; Sviben, Igor ; Rokov Plavec, Jasmina (ur.).
Zagreb: Faculty of Science, University of Zagreb, 2015. str. 22-22 (poster, nije recenziran, sažetak, znanstveni)


CROSBI ID: 767751 Za ispravke kontaktirajte CROSBI podršku putem web obrasca

Naslov
Plant seryl-tRNA synthetase and metabolic protein BEN1: identification of interaction surfaces by biophysical methods

Autori
Kekez, Mario ; Zanki, Vladimir ; Hodnik, Vesna ; Anderluh, Gregor ; Rokov Plavec, Jasmina

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Izvornik
Symposium & Workshop on Microscale Thermophoresis: Book of Abstracts / Godinić Mikulčić, Vlatka ; Sviben, Igor ; Rokov Plavec, Jasmina - Zagreb : Faculty of Science, University of Zagreb, 2015, 22-22

ISBN
978-953-6076-37-6

Skup
Symposium & Workshop on Microscale Thermophoresis

Mjesto i datum
Zagreb, Hrvatska, 30.06-01.07.2015

Vrsta sudjelovanja
Poster

Vrsta recenzije
Nije recenziran

Ključne riječi
protein-protein interaction ; BEN1 ; plant seryl-tRNA synthetase ; microscale thermophoresis ; surface plasmon resonance

Sažetak
Aminoacyl-tRNA synthetases (aaRS) attach appropriate amino acids to cognate tRNAs ensuring efficient protein biosynthesis. Their involvement in diverse celullar functions beyond translation opens broad new perspectives in functional proteomics, especially in the field of plant aaRSs which is fairly uninvestigated. To shed light on non-canonical functions of cytosolic seryl-tRNA synthetase (SerRS) from plant Arabidopsis thaliana we conducted yeast two hybrid screen (Y2H) which revealed metabolic protein BEN1 as a highly promising interacting partner. Interaction was further biophysicaly analyzed in vitro using isothermal calorimetry titration (ITC), pull- down, surface plasmon resonance (SPR) and microscale thermophoresis method (MST). Due to the nature of interaction and sensitivity of applied methods we were not able to retrieve positive results using pull-down assay and ITC, but SPR and MST gave us postive confirmation of interaction and information about dissociation constant (Kd constant obtained using SPR = 1, 03 × 10-6 (± 1, 67 × 10-7) mol dm-3, in good agreement with Kd obtained using MST = 4, 45 × 10-7 (± 1, 40 × 10-7) mol dm-3). To determine interaction surfaces and pinpoint regions responsible for SerRS and BEN1 interaction, truncated variants of both SerRS and BEN1 proteins were prepared and protein interactions were analyzed using MST. Kd for complex containing BEN1 and truncated SerRS variant without basic C-terminal extension (with or without (His)6 tag) was similar to Kd of the SerRS:BEN1 complex indicating that SerRS basic C- terminal extension does not have any influence on interaction. Furthermore, isolated N-terminal domain of SerRS did not form the complex with BEN1. Taking into account observed data, we concluded that interaction between SerRS and BEN1 involves the central part of SerRS that contains globular catalytic domain. Determining interaction domain of BEN1 was not possible because of aggregation problems in MST assay when using truncated BEN1 version, so this part of investigation is yet to be done. Knowing precise interaction contacts will allow us to understand and propose functional importance of SerRS:BEN1 assembly in plant organism.

Izvorni jezik
Engleski

Znanstvena područja
Kemija, Biologija



POVEZANOST RADA


Projekti:
MZOS-119-0982913-1358 - Strukturna raznolikost seril-tRNA sintetaza i točnost biosinteze proteina (Rokov Plavec, Jasmina; Weygand Đurašević, Ivana, MZOS ) ( POIROT)

Ustanove:
Prirodoslovno-matematički fakultet, Zagreb

Profili:

Avatar Url Vladimir Zanki (autor)

Avatar Url Jasmina Rokov Plavec (autor)

Avatar Url Mario Kekez (autor)


Citiraj ovu publikaciju:

Kekez, Mario; Zanki, Vladimir; Hodnik, Vesna; Anderluh, Gregor; Rokov Plavec, Jasmina
Plant seryl-tRNA synthetase and metabolic protein BEN1: identification of interaction surfaces by biophysical methods // Symposium & Workshop on Microscale Thermophoresis: Book of Abstracts / Godinić Mikulčić, Vlatka ; Sviben, Igor ; Rokov Plavec, Jasmina (ur.).
Zagreb: Faculty of Science, University of Zagreb, 2015. str. 22-22 (poster, nije recenziran, sažetak, znanstveni)
Kekez, M., Zanki, V., Hodnik, V., Anderluh, G. & Rokov Plavec, J. (2015) Plant seryl-tRNA synthetase and metabolic protein BEN1: identification of interaction surfaces by biophysical methods. U: Godinić Mikulčić, V., Sviben, I. & Rokov Plavec, J. (ur.)Symposium & Workshop on Microscale Thermophoresis: Book of Abstracts.
@article{article, author = {Kekez, Mario and Zanki, Vladimir and Hodnik, Vesna and Anderluh, Gregor and Rokov Plavec, Jasmina}, year = {2015}, pages = {22-22}, keywords = {protein-protein interaction, BEN1, plant seryl-tRNA synthetase, microscale thermophoresis, surface plasmon resonance}, isbn = {978-953-6076-37-6}, title = {Plant seryl-tRNA synthetase and metabolic protein BEN1: identification of interaction surfaces by biophysical methods}, keyword = {protein-protein interaction, BEN1, plant seryl-tRNA synthetase, microscale thermophoresis, surface plasmon resonance}, publisher = {Faculty of Science, University of Zagreb}, publisherplace = {Zagreb, Hrvatska} }
@article{article, author = {Kekez, Mario and Zanki, Vladimir and Hodnik, Vesna and Anderluh, Gregor and Rokov Plavec, Jasmina}, year = {2015}, pages = {22-22}, keywords = {protein-protein interaction, BEN1, plant seryl-tRNA synthetase, microscale thermophoresis, surface plasmon resonance}, isbn = {978-953-6076-37-6}, title = {Plant seryl-tRNA synthetase and metabolic protein BEN1: identification of interaction surfaces by biophysical methods}, keyword = {protein-protein interaction, BEN1, plant seryl-tRNA synthetase, microscale thermophoresis, surface plasmon resonance}, publisher = {Faculty of Science, University of Zagreb}, publisherplace = {Zagreb, Hrvatska} }




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