Naslov
Identification of protein-protein interactions in snake venom by immuno-affinity chromatography on monoliths

Autori
Brgles, Marija ; Kurtović, Tihana ; Kovačič, Lidija ; Križaj, Igor ; Barut, Miloš ; Lang Balija, Maja ; Allmaier, Günter ; Marchetti-Deschmann, Martina ; Halassy, Beata

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

Izvornik
Book of Abstracts 6th Monolith Summer School & Symposium / - , 2014, 39-39

Skup
6th Monolith Summer School & Symposium

Mjesto i datum
Portorož, Slovenija, 30.05.2014. - 04.06.2014

Vrsta sudjelovanja
Predavanje

Vrsta recenzije
Međunarodna recenzija

Ključne riječi
snake venom; protein-protein interactions; immuno-affinity chromatography

Sažetak
In order to perform their function, proteins frequently interact with other proteins. Snake venom is composed mostly of proteins, and various protein complexes in the venom have been found to exhibit higher toxicity levels than respective components separately. Complexes can modulate envenomation activity of a venom and/or potentiate its effect. Our previous data indicate that the most toxic components of the Vipera ammodytes ammodytes (Vaa) venom isolated so far – ammodytoxins (Atxs) are contributing to the venom’s toxicity only moderately, therefore we aimed to explore whether they have some interacting partner(s) potentiating toxicity. Various methods are used to reveal protein interacting partners and affinity chromatography is one of them. For screening of possible interactions in Vaa venom immuno-affinity chromatography combined with identification by mass spectrometry was used. Various chemistries (epoxy, carbonyldiimidazole, ethylenediamine) as well as protein G functionality were used to immobilize anti-Atx antibodies on monolith support, a Convective Interaction Media disk. Using such approach several proteins were indicated as potential Atx-binding proteins. Among these, the interaction of Atxs with a Kunitz-type inhibitor was confirmed by far-Western dot-blot and surface plasmon resonance measurement. It can be concluded that affinity chromatography on monoliths combined with mass spectrometry identification is a successful approach for screening of protein interactions and it resulted with detection of the interaction of Atx with Kunitz-type inhibitor in Vaa venom for the first time. (M. Brgles, T. Kurtović, L. Kovačić, I. Križaj, M. Barut, M. Lang Balija, G. Allmaier, M. Marchetti-Deschmann, B. Halassy, Anal. Bioanal. Chem. 406 (2014) 293-304.)

Izvorni jezik
Engleski

Znanstvena područja
Kemija, Biotehnologija

POVEZANOST RADA