Phenotypic selection and characterization of randomly produced non-haemolytic mutants of the toxic sea anemone protein sticholysin II

FEBS Letters
Jorge Alegre-CebolladaA Martínez del Pozo

Abstract

A rapid screening method for haemolytic activity, using blood agar plates, has been developed to analyze randomly produced mutant variants of the pore-forming protein sticholysin II (Stn II). Those exhibiting a reduced activity were selected and the DNA corresponding to each Stn II variant sequenced. Once the mutation produced was determined, protein variants were isolated and characterized in terms of structure (circular dichroism spectra and thermal stability) and haemolytic activity. Three single mutation protein variants, at residues K19, F106 and Y111, showed a significantly decreased haemolytic activity while their thermostability was identical to that of the wild-type protein. Considering the obtained data and based on the three-dimensional structure of the protein, the role of these residues on the mechanism of haemolysis has been analyzed.

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Citations

May 14, 2010·The Journal of Biological Chemistry·Biserka BakracGregor Anderluh
Feb 5, 2013·Archives of Biochemistry and Biophysics·Sara García-LinaresJosé G Gavilanes
Sep 10, 2015·Biochimica Et Biophysica Acta·Nejc RojkoGregor Anderluh
Aug 10, 2010·Toxicon : Official Journal of the International Society on Toxinology·Margarita MonastyrnayaEmma Kozlovskaya
Mar 10, 2009·Toxicon : Official Journal of the International Society on Toxinology·Katarina Crnigoj KristanGregor Anderluh
Mar 10, 2009·Toxicon : Official Journal of the International Society on Toxinology·Carlos AlvarezMaría E Lanio
Apr 22, 2010·Proteins·Inés CastrilloMarta Bruix
Oct 28, 2006·Toxicon : Official Journal of the International Society on Toxinology·F PazosE Lissi
May 31, 2011·Biochimica Et Biophysica Acta·Lucía García-OrtegaJosé G Gavilanes
Apr 23, 2011·Toxicon : Official Journal of the International Society on Toxinology·A ValleS Schreier
Aug 9, 2008·Journal of Molecular Biology·Jorge Alegre-CebolladaAlvaro Martínez-del-Pozo
Jun 19, 2007·Biophysical Journal·Jorge Alegre-CebolladaErik Goormaghtigh
Jan 24, 2018·Biopolymers·Aline Lima de OliveiraAlberto Spisni
Apr 30, 2016·The Journal of Biological Chemistry·Esperanza Rivera-de-TorreÁlvaro Martínez-Del-Pozo
Apr 30, 2008·The Journal of Biological Chemistry·Biserka BakracGregor Anderluh
Dec 2, 2020·International Journal of Molecular Sciences·Esperanza Rivera-de-TorreSara García-Linares
Jun 3, 2020·Archives of Biochemistry and Biophysics·Esperanza Rivera-de-TorreÁlvaro Martínez-Del-Pozo
Jul 12, 2021·Biochimica Et Biophysica Acta. Proteins and Proteomics·Juan Palacios-OrtegaÁlvaro Martínez-Del-Pozo

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