Abstract
The recent global challenges to prevent and treat fungal infections strongly demand for the development of new antifungal strategies. The structurally very similar cysteine-rich antifungal proteins from ascomycetes provide a feasible basis for designing new antifungal molecules. The main structural elements responsible for folding, stability and antifungal activity are not fully understood, although this is an essential prerequisite for rational protein design. In this study, we used the Neosartorya fischeri antifungal protein (NFAP) to investigate the role of the disulphide bridges, the hydrophobic core, and the N-terminal amino acids in the formation of a highly stable, folded, and antifungal active protein. NFAP and its mutants carrying cysteine deletion (NFAPΔC), hydrophobic core deletion (NFAPΔh), and N-terminal amino acids exchanges (NFAPΔN) were produced in Pichia pastoris. The recombinant NFAP showed the same features in structure, folding, stability and activity as the native protein. The data acquired with mass spectrometry, structural analyses and antifungal activity assays of NFAP and its mutants proved the importance of the disulphide bonding, the hydrophobic core and the correct N-terminus for folding, stability a...Continue Reading
References
Mar 7, 1995·Biochemistry·R Campos-OlivasM Rico
Dec 20, 1995·Archives of Biochemistry and Biophysics·J LacadenaJ G Gavilanes
Nov 1, 1995·Proteins·A SaliM Karplus
Jun 20, 1997·Biochimica Et Biophysica Acta·A Martínez-RuizJ G Gavilanes
Dec 9, 2000·Analytical Biochemistry·N Sreerama, R W Woody
Aug 24, 2001·Journal of Molecular Biology·C W CarterM H Edgell
Jan 24, 2002·Proceedings of the National Academy of Sciences of the United States of America·Margaret S CheungJosé N Onuchic
Jan 31, 2003·Proteins·Simon C LovellDavid C Richardson
Jul 21, 2004·Journal of Computational Chemistry·Eric F PettersenThomas E Ferrin
Dec 4, 2004·Protein Engineering, Design & Selection : PEDS·Rajasri BhattacharyyaPinak Chakrabarti
Dec 8, 2004·Journal of Leukocyte Biology·Hans-Georg SahlAlessandro Tossi
Feb 24, 2006·Journal of Peptide Science : an Official Publication of the European Peptide Society·Enno KlüverAxel Schulz
Jun 22, 2006·Bioinformatics·Jonathan G LeesB A Wallace
Jul 18, 2006·Nucleic Acids Research·Alessio CeroniPaolo Frasconi
Apr 5, 2007·Nature Protocols·Norma J Greenfield
Oct 30, 2007·Cellular and Molecular Life Sciences : CMLS·F MarxI Pócsi
Nov 28, 2007·Biopolymers·Karen TaylorJulia R Dorin
Dec 11, 2007·Applied Microbiology and Biotechnology·Vera Meyer
Jan 25, 2008·Molecular Biology of the Cell·Naimeh Taheri-TaleshBerl R Oakley
May 23, 2009·The FEBS Journal·Gyula BattaFlorentine Marx
Mar 24, 2010·Antimicrobial Agents and Chemotherapy·Olga ScudieroFrancesco Salvatore
Jul 12, 2011·Peptides·Laura KovácsLászló Galgóczy
Sep 13, 2011·Peptides·Marcus SeiboldOle Olsen
Nov 22, 2011·Biophysical Journal·Dong Xu, Yang Zhang
Dec 1, 2012·Microbiology·László GalgóczyCsaba Vágvölgyi
Nov 1, 2013·Chemistry : a European Journal·Györgyi VáradiGyula Batta
Nov 26, 2013·Protein Expression and Purification·Máté VirághLászló Galgóczy
Apr 30, 2014·Molecular Microbiology·Alberto MuñozNick D Read
Feb 14, 2015·Chemistry : a European Journal·Ádám FizilGyula Batta
May 23, 2015·Protein & Cell·Máté VirághLászló Galgóczy
Jul 11, 2016·Applied Microbiology and Biotechnology·Josué DelgadoFélix Núñez
Jul 28, 2016·Scientific Reports·Carlo CamilloniMichele Vendruscolo
Nov 12, 2016·Microbial Cell Factories·Christoph SondereggerFlorentine Marx
Jan 11, 2017·PloS One·Christoph SondereggerFlorentine Marx
Sep 29, 2017·Fungal biology and biotechnology·Vera MeyerRitchie M Head
Citations
Nov 28, 2018·Antimicrobial Agents and Chemotherapy·Renátó KovácsLászló Galgóczy
Jan 31, 2018·Scientific Reports·Anna HuberGyula Batta
Mar 25, 2020·Microbial Biotechnology·Liliána TóthLászló Galgóczy
Jun 2, 2018·Oxidative Medicine and Cellular Longevity·Michael R YeamanKarin Thevissen
Nov 9, 2017·Scientific Reports·Sandra GarriguesPaloma Manzanares
Nov 5, 2019·Israel Journal of Chemistry·László GalgóczyFlorentine Marx
Aug 23, 2020·Journal of Fungi·Jeanett HolzknechtFlorentine Marx
Apr 24, 2020·PLoS Pathogens·Sheng TongYanhua Fan
Mar 23, 2018·Frontiers in Microbiology·Liliána TóthLászló Galgóczy
Jun 2, 2020·Frontiers in Microbiology·Liliána TóthLászló Galgóczy
Jan 29, 2021·International Journal of Molecular Sciences·András CzajlikGyula Batta
Mar 4, 2021·Food Microbiology·Pedro V Martínez-CulebrasPaloma Manzanares