Structural determinants of Neosartorya fischeri antifungal protein (NFAP) for folding, stability and antifungal activity

Scientific Reports
László GalgóczyFlorentine Marx

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

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
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
Jul 18, 2006·Nucleic Acids Research·Alessio CeroniPaolo Frasconi
Apr 5, 2007·Nature Protocols·Norma J Greenfield
Nov 28, 2007·Biopolymers·Karen TaylorJulia R Dorin
Jan 25, 2008·Molecular Biology of the Cell·Naimeh Taheri-TaleshBerl R Oakley
Mar 24, 2010·Antimicrobial Agents and Chemotherapy·Olga ScudieroFrancesco Salvatore
Jul 12, 2011·Peptides·Laura KovácsLászló Galgóczy
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

❮ Previous
Next ❯

Citations

Nov 28, 2018·Antimicrobial Agents and Chemotherapy·Renátó KovácsLászló Galgóczy
Jun 2, 2018·Oxidative Medicine and Cellular Longevity·Michael R YeamanKarin Thevissen
Jan 29, 2021·International Journal of Molecular Sciences·András CzajlikGyula Batta
Mar 4, 2021·Food Microbiology·Pedro V Martínez-CulebrasPaloma Manzanares

❮ Previous
Next ❯

Methods Mentioned

BETA
electrophoresis
protein folding
nuclear magnetic resonance
circular dichroism
fluorescence microscopy

Software Mentioned

MODELLER
ExPASy ProtParam tool Calculator
DISULFIND Cysteines Disulfide Bonding State Connectivity Predi...
UCSF Chimera
GraphPad Prism
GraphPad
Excel
Xcalibur pXtract
RAMPAGE
ModRefiner

Related Concepts

Related Feeds

Antifungals (ASM)

An antifungal, also known as an antimycotic medication, is a pharmaceutical fungicide or fungistatic used to treat and prevent mycosis such as athlete's foot, ringworm, candidiasis, cryptococcal meningitis, and others. Discover the latest research on antifungals here.

Antifungals

An antifungal, also known as an antimycotic medication, is a pharmaceutical fungicide or fungistatic used to treat and prevent mycosis such as athlete's foot, ringworm, candidiasis, cryptococcal meningitis, and others. Discover the latest research on antifungals here.