Naslov
The TROL protein: one protein approach and the consequences

Autori
Jurić, Snježana ; Tomašić, Ana ; Hazler-Pilepić, Kroata ; Lepeduš, Hrvoje ; Jeličić, Branka ; Fulgosi, Hrvoje

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

Izvornik
Tenth International Summer School on Biophysics: "Supramolecular Structure and Function" / Pifat-Mrzljak, Greta ; Zahradka, Ksenija - Zagreb : Institut Ruđer Bošković, 2009, 119-119

ISBN
978-953-6690-81-7

Skup
Tenth International Summer School on Biophysics: "Supramolecular Structure and Function"

Mjesto i datum
Rovinj, Hrvatska, 19.9.09.-01.10.09

Vrsta sudjelovanja
Poster

Vrsta recenzije
Domaća recenzija

Ključne riječi
Photosynthesis; Rhodanese; Signalling

Sažetak
Our current research is focused on defining the role of the thylakoid membrane proteins in Arabidopsis plants. By using standard molecular biology methods and procedures, we were able to answer the questions about the role of the protein entitled TROL (thylakoid rhodanese-like protein). Localization studies using C-terminal YFP fusion revealed chloroplast localization of TROL. In vitro chloroplast import assays confirmed its thylakoid localization and have demonstrated that TROL possesses characteristics of an integral membrane component. Furthermore, TROL is most likely exclusively located in non-apressed thylakoid regions. A strong interaction between TROL and FNR was demonstrated by co-immunoprecipitation experiments and yeast two hybrid methods. TROL possesses a rhodanese-like domain linked to a module which interacts with FNR. Rhodaneses are very old group of proteins, suggested to be involved in distinct biological functions such as cyanide detoxification, formation of prosthetic groups in iron-sulfur proteins and thiamin biosynthesis. Our aim is to produce transgenic plants with modified protein TROL to explore the function of the rhodanese – like domain. Analysis of Arabidopsis mutant lines indicates that, in the absence of TROL, relative electron transport rates at high-light intensities are severely lowered accompanied with significant increase in non-photochemical quenching (NPQ). I have chosen this title because during my first five years at RBI I have learned many techniques by exploring only one protein, and how much effort is put into discovering its role in vivo. I started with growing Arabidopsis plants and transforming them, then moved forward to exploring protein-protein interactions only to discover that I needed to find out more about the environment of my protein TROL, about the interacting partners, the complexes it is possibly involved in. By using blue-native polyacrilamide gel electrophoresis technique (BN-PAGE), we were able to show that TROL could be found in the three major complexes, at 110 kDa, 120 kDa, and 420 kDa. The 420 kDa complex might contain representative subunits of cytochrome b6/f, PSI, and PSII. TROL associated with FNR could be found in a minor, probably transient, complex at about 190 kDa. I would like to present the techniques we are currently using in my laboratory and to get a feedback on how to continue in the field of exploring protein complexes.

Izvorni jezik
Engleski

Znanstvena područja
Biologija

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