Recently, we documented that the short, proline-rich antibacterial peptides pyrrhocoricin, drosocin, and apidaecin interact with bacterial heat shock protein DnaK, peptide binding to DnaK can be correlated antimicrobial activity. In current report studied mechanism of action these their sites Escherichia coli DnaK. Biologically active pyrrhocoricin made l-amino acids diminished ATPase activity recombinant The inactive d-pyrrhocoricin analogue membrane-active cecropin A or magainin 2 failed...

The increasing incidence of fungal infections and damages due to drug-resistant fungi urges the development new antifungal strategies. cysteine-rich proteins from filamentous ascomycetes provide a feasible base for protection against molds their potent activity on them. In contrast this, they show no or weak yeasts, hence applicability this group is questionable. present study 5.6 kDa anti-yeast protein (NFAP2) isolated, identified characterized ferment broth Neosartorya fischeri NRRL 181....

In response to the growth of emerging resistance conventional antifungal drugs, proteins (AFPs) filamentous Ascomycetes origin have been discovered in recent years. Understanding structure AFPs is crucial for elucidating their mechanisms and developing new therapeutic agents. While nuclear magnetic resonance (NMR) has proven effective determining structures small proteins, some AFP remain unresolved, necessitating use alternative prediction methods. Through bioinformatics analysis heatmaps...

The increasing number of life-threatening Candida infections caused by antifungal drug-resistant strains urges the development new therapeutic strategies. small, cysteine-rich, and cationic Neosartorya fischeri protein 2 (NFAP2) effectively inhibits growth spp. Limiting factors its future application, are low-yield production native producer, unavailable information about potential clinical unsolved relationship between structure function. In present study we adopted a Penicillium...

Candida auris is a potential multidrug-resistant pathogen able to persist on indwelling devices as biofilm, which serve source of catheter-associated infections. Neosartorya fischeri antifungal protein 2 (NFAP2) cysteine-rich, cationic with potent anti-Candida activity. We studied the in vitro activity NFAP2 alone and combination fluconazole, amphotericin B, anidulafungin, caspofungin, micafungin against C. biofilms. The nature interactions was assessed utilizing fractional inhibitory...

Miniproteins are adequate models to study various protein-structure modifying effects such as temperature, pH, point mutation(s), H-bonds, salt bridges, molecular packing, etc. Tc5b, a 20-residue Trp-cage protein is one of the smallest with stable 3D fold (Neidigh J. W. et al. (2002) Nat. Struct. Biol. 9, 425−430). However, Tc5b exhibits considerable heat-sensitivity and only at relatively low temperatures. Here we report systematic investigation structural factors influencing stability by...

Small, cysteine-rich and cationic antimicrobial proteins (AMPs) from filamentous ascomycetes represent ideal bio-molecules for the development of next-generation antifungal therapeutics. They are promising candidates to counteract resistance may complement or even replace current small molecule-based antibiotics in future. In this study, we show that a 14 amino acid (aa) long peptide (Pγ) spanning highly conserved γ-core motif Penicillium chrysogenum protein (PAF) has activity against...

As a consequence of emerging numbers vulvovaginitis cases caused by azole-resistant and biofilm-forming Candida species, fast efficient treatment this infection has become challenging. The problem is further exacerbated the severe side effects azoles as long-term-use medications in recurrent form. There therefore an increasing demand for novel safely applicable effective antifungal therapeutic strategies. small, cysteine-rich, cationic proteins from filamentous ascomycetes are potential...

Small, cysteine-rich and cationic antimicrobial proteins (AMPs) from filamentous ascomycetes promise treatment alternatives to licensed antifungal drugs. In this study, we characterized the Penicillium chrysogenum Q176 protein C (PAFC), which is phylogenetically distinct other two proteins, PAF PAFB, that are expressed by biotechnologically important ascomycete. PAFC secreted into culture broth co-expressed with PAFB in exudates of surface cultures. This observation line suggested role AMPs...

Plant pathogenic fungi are responsible for enormous crop losses worldwide. Overcoming this problem is challenging as these can be highly resistant to approved chemical fungicides. There thus a need develop and introduce fundamentally new plant protection strategies sustainable agricultural production. Highly stable extracellular antifungal proteins (AFPs) their rationally designed peptide derivatives (PDs) constitute feasible options meet challenge. In the present study, potential topical...

Summary The prevention of enormous crop losses caused by pesticide‐resistant fungi is a serious challenge in agriculture. Application alternative fungicides, such as antifungal proteins and peptides, provides promising basis to overcome this problem; however, their direct use fields suffers limitations, high cost production, low stability, narrow spectrum toxicity on plant or mammalian cells. Recently, we demonstrated that Penicillium chrysogenum ‐based expression system feasible tool for...

Abstract The folding of disulfide proteins is considerable interest because knowledge this may influence our present understanding protein folding. However, sometimes even the pattern cannot be unequivocally determined by available experimental techniques. For example, structures a few small antifungal (PAF, AFP) have been disclosed recently using NMR spectroscopy but with some ambiguity in actual pattern. reason, we carried out chemical synthesis PAF. Probing different approaches, oxidative...

The recent global challenges to prevent and treat fungal infections strongly demand for the development of new antifungal strategies. structurally very similar cysteine-rich proteins from ascomycetes provide a feasible basis designing molecules. main structural elements responsible folding, stability activity are not fully understood, although this is an essential prerequisite rational protein design. In study, we used Neosartorya fischeri (NFAP) investigate role disulphide bridges,...

Because of enormous crop losses worldwide due to pesticide-resistant plant pathogenic fungi, there is an increasing demand for the development novel antifungal strategies in agriculture. Antifungal proteins and peptides are considered potential biofungicides; however, several factors limit their direct agricultural application, such as high cost production, narrow spectrum detrimental effects human/animal health. This study evaluated safety application protein from ascomycete Neosartorya...

Antifungal peptides offer promising alternative compounds for the treatment of fungal infections, which new antifungal are urgently needed. Constant and broad spectra these play essential roles in their reliable therapeutic application. It has been observed that rationally designed using evolutionarily conserved γ-core region (GXC–X3–9–C) an protein from Neosartorya (Aspergillus) fischeri highly inhibit growth fungi. The cysteines have free sulfhydryl groups, allow cyclization dimerization...

The 20 residue long Trp-cage miniprotein is an excellent model for both computational and experimental studies of protein folding stability. Recently, great attention emerged to study disease-related misfolding, aggregation, amyloid formation, with the aim revealing their structural thermodynamic background. sensitive environmental structure-modifying effects. It aggregates ease upon structure destabilization, thus it suitable modeling aggregation formation. Here, we characterize formation...

As a consequence of the fast resistance spreading, limited number drugs are available to treat fungal infections. Therefore, there is an urgent need develop new antifungal treatment strategies. The features disulfide bond-stabilized protein, NFAP2 secreted by mold Neosartorya (Aspergillus) fischeri render it be promising template for future protein-based drug design, which requires knowledge about native linkage pattern as one prerequisites biological activity. However, in lack tryptic and...

The genome of Penicillium chrysogenum Q176 contains a gene coding for the 88-amino-acid (aa)-long glycine- and cysteine-rich P. antifungal protein C (PAFC). After maturation, secreted miniprotein (MP) comprises 64 aa shares 80% identity with bubble (BP) from brevicompactum, which has published X-ray structure. Our team expressed isotope (15N, 13C)-labeled, recombinant PAFC in high yields, allowed us to determine solution structure molecular dynamics by nuclear magnetic resonance (NMR)...

Emerging fungal infections require new, more efficient antifungal agents and therapies. AFP, a protein from Aspergillus giganteus with four disulfide bonds, is promising candidate because it selectively inhibits the growth of filamentous fungi. In this work, reduced form AFP was prepared using native chemical ligation. The synthesized via oxidative folding uniform protection for cysteine thiols. AFP's biological activity depends heavily on pattern natural bonds. Enzymatic digestion MS...

The influenza virus haemagglutinin has an important role in the infectious cycle of and carries multiple B T cell epitopes. It is synthesized as a single polypeptide chain but viral infectivity depends on its post‐translational enzymatic cleavage. cleavage site trypsin‐like enzyme responsible for this modification found most conserved intersubunit region molecule. In study antibody recognition was investigated. Synthetic peptides comprising intact cleaved forms segment were used to examine...