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    Functional characterization and FTIR-based 3D modeling of full length and truncated forms of Scorpio maurus venom phospholipase A(2)

    Year: 2018

    Journal: Biochim. Biophys. Acta-Gen. Subj., Volume 1862, MAY, page 1247–1261

    Authors: Krayem, Najeh; Parsiegla, Goetz; Gaussier, Helene; Louati, Hanen; Jallouli, Raida; Mansuelle, Pascal; Carriere, Frederic; Gargouri, Youssef

    Organizations: Ministry of Higher Education, Scientific Research in Tunisia [1894/2015]

    Keywords: Secreted phospholipase A(2); Heterologous expression; Enzyme activity; Molecular modeling; FTTR spectroscopy

    Background: Heterodimeric phospholipase A(2) from venom glands of Tunisian scorpion Scorpio maurus (Sm-PLGV) had been purified. It contains long and short chains linked by a disulfide bridge. Sm-PLGV exhibits hemolytic activity towards human erythrocytes and interacts with phospholipid monolayers at high surface pressure. The investigation of structure-function relationships should provide new clues to understand its activity. Methods: Molecular cloning of Sm-PLGV and heterologous expression in Escherichia coli of three recombinant forms was used to determine the role of the short chain on enzymatic activity. Infrared spectroscopy assisted 3D model building of the three recombinant constructs (phospholipases with and without the penta-peptide and Long chain only) allowed us to propose an explanation of the differences in specific activities and their interaction with various phospholipids. Results: Nucleotide sequence of Sm-PLGV encodes 129 residues corresponding to the Long chain, the penta-peptide and the short chain. Although recombinant phospholipases without and with the penta-peptide have different specific activities, they display a similar substrate specificity on various phospholipid monolayers and similar bell-shaped activity profiles with maxima at high surface pressure. The absence of the short chain reduces significantly enzymatic and hemolytic activities. The 3D models pointed to an interaction of the short chain with the catalytic residues, what might explain the difference in activities of our constructs. Conclusion: Infrared spectroscopy data and 3D modeling confirm the experimental findings that highlight the importance of the short chain for the Sm-PLGV activity.
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