Naslov
Isolation of ammodytoxin A from Vipera ammodytes ammodytes venom using two step chromatography

Autori
Kurtović, Tihana ; Brgles, Marija ; Marchetti-Deschmann, Martina ; Križaj, Igor ; Barut, Miloš ; Allmaier, Günter ; Halassy, Beata

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

Izvornik
Book of Abstracts, MSS2010-4rd Monolith Summer School and Symposium / Štrancar, Aleš - , 2010, 36-36

Skup
MSS2010-4rd Monolith Summer School and Symposium, Applications in Biochromatography, Bioconversion and Solid Phase Synthesis

Mjesto i datum
Portorož, Slovenija, 30.05.2010. - 02.06.2010

Vrsta sudjelovanja
Poster

Vrsta recenzije
Međunarodna recenzija

Ključne riječi
Ammodytoxins; CIM disk; Mass spectrometry

Sažetak
Ammodytoxins (Atxs) are phospholipases A2 acting as presynaptic neurotoxins. Three isoforms (A, B and C) have been identified in Vipera ammodytes ammodytes venom. The amount of Atxs in the venom correlates with venom's toxicity and immunogenicity. AtxA (the most toxic isoform) differs from AtxB in three and from AtxC in two amino acid residues, resulting in 2 and 33 Da mass differences. Isoelectric points of AtxA, AtxB and AtxC are 10.2, 10.0 and 9.4, respectively. Chromatography of pure Atx isoforms on CM-CIM disk under alkaline conditions resulted in separate elution of AtxA from AtxB and AtxC. We hypothesized that this allows the isolation of pure AtxA from crude venom so cation exchange chromatography of crude venom combined with affinity chromatography was performed, but this procedure did not result in isolation of pure AtxA. Mass spectrometry characterization (molecular mass determination of intact proteins, peptide mass fingerprinting and tandem mass spectrometry for sequence determination) of purified Atx fractions enabled identification of all three isoforms.

Izvorni jezik
Engleski

Znanstvena područja
Kemija, Biologija, Temeljne medicinske znanosti

POVEZANOST RADA