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1.

001-es BibID:BIBFORM008914
Első szerző:Batta Gyula (molekula-szerkezet kutató)
Cím:Functional aspects of the solution structure and dynamics of PAF - a highly-stable antifungal protein from Penicillium chrysogenum / Gyula Batta, Teréz Barna, Zoltán Gáspári, Szabolcs Sándor, Katalin E. Kövér, Ulrike Binder, Bettina Sarg, Lydia Kaiserer, Anil Kumar Chhillar, Andrea Eigentler, Éva Leiter, Nikoletta Hegedűs, István Pócsi, Herbert Lindner, Florentine Marx
Dátum:2009
Tárgyszavak:Természettudományok Kémiai tudományok idegen nyelvű folyóiratközlemény külföldi lapban
Molekulatudomány
antifungal protein PAF
internal dynamics
NMR spectroscopy
site-directed
mutagenesis
solution structure
Megjelenés:The FEBS Journal. - 276 : 10 (2009), p. 2875-2890. -
További szerzők:Barna Teréz (1963-) (vegyész) Gáspári Zoltán Sándor Szabolcs Kövér Katalin, E. (1956-2023) (vegyész) Binder, Ulrike Sarg, Bettina Kaiserer, Lydia Chhillar, Anil Kumar Eigentler, Andrea Leiter Éva (1976-) (biológus) Hegedűs Nikoletta Pócsi István (1961-) (vegyész) Lindner, Herbert Marx, Florentine
Pályázati támogatás:TÁMOP-4.2.2-08/1-2008-0019
TÁMOP
Internet cím:DOI
elektronikus változat
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2.

001-es BibID:BIBFORM058875
Első szerző:Binder, Ulrike
Cím:Protein kinase A signaling and calcium ions are major players in PAF mediated toxicity against Aspergillus niger / Ulrike Binder, Mojca Benčina, Ádám Fizil, Gyula Batta, Anil K. Chhillar, Florentine Marx
Dátum:2015
ISSN:0014-5793 1873-3468
Megjegyzések:The Penicillium chrysogenum antifungal protein PAF is toxic against potentially pathogenic Ascomycetes. We used the highly sensitive aequorin-expressing model Aspergillus niger to identify a defined change in cytoplasmic free Ca2+ dynamics in response to PAF. This Ca2+ signature depended on an intact positively charged lysine-rich PAF motif. By combining Ca2+ measurements in A. niger mutants with deregulated cAMP/protein kinase A (PKA) signaling, we proved the interconnection of Ca2+ perturbation and cAMP/PKA signaling in the mechanistic function of PAF. A deep understanding of the mode of action of PAF is an invaluable prerequisite for its future application as new antifungal drug.
Tárgyszavak:Természettudományok Kémiai tudományok idegen nyelvű folyóiratközlemény külföldi lapban
Antifungal protein
PAF
Aspergillus niger
Calcium homeostasis
Protein kinase A signaling
Megjelenés:Febs Letters. - 589 : 11 (2015), p. 1266-1271. -
További szerzők:Bencina, Mojca Fizil Ádám (1988-) (biológus) Batta Gyula (1953-) (molekula-szerkezet kutató) Chhillar, Anil Kumar Marx, Florentine
Pályázati támogatás:ANN 110821
OTKA
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3.

001-es BibID:BIBFORM090877
035-os BibID:(cikkazonosító)1183 (scopus)85099978064 (wos)000615359000001
Első szerző:Czajlik András (gyógyszerész)
Cím:Solution Structure, Dynamics, and New Antifungal Aspects of the Cysteine-Rich Miniprotein PAFC / András Czajlik, Jeanett Holzknecht, László Galgóczy, Liliána Tóth, Péter Poór, Attila Ördög, Györgyi Váradi, Alexander Kühbacher, Attila Borics, Gábor K. Tóth, Florentine Marx, Gyula Batta
Dátum:2021
ISSN:1661-6596 1422-0067
Megjegyzések:The genome of Penicillium chrysogenum Q176 contains a gene coding for the 88-amino-acid (aa)-long glycine- and cysteine-rich P. chrysogenum antifungal protein C (PAFC). After maturation, the secreted antifungal miniprotein (MP) comprises 64 aa and shares 80% aa identity with the bubble protein (BP) from Penicillium brevicompactum, which has a published X-ray structure. Our team expressed isotope (15N, 13C)-labeled, recombinant PAFC in high yields, which allowed us to determine the solution structure and molecular dynamics by nuclear magnetic resonance (NMR) experiments. The primary structure of PAFC is dominated by 14 glycines, and therefore, whether the four disulfide bonds can stabilize the fold is challenging. Indeed, unlike the few published solution structures of other antifungal MPs from filamentous ascomycetes, the NMR data indicate that PAFC has shorter secondary structure elements and lacks the typical ??-barrel structure, though it has a positively charged cavity and a hydrophobic core around the disulfide bonds. Some parts within the two putative ??-core motifs exhibited enhanced dynamics according to a new disorder index presentation of 15N-NMR relaxation data. Furthermore, we also provided a more detailed insight into the antifungal spectrum of PAFC, with specific emphasis on fungal plant pathogens. Our results suggest that PAFC could be an effective candidate for the development of new antifungal strategies in agriculture.
Tárgyszavak:Természettudományok Kémiai tudományok idegen nyelvű folyóiratközlemény külföldi lapban
folyóiratcikk
Penicillium chrysogenum
antifungal protein PAFC
γ-core motif
solution structure
dynamics
nuclear magnetic resonance
plant protection
Megjelenés:International Journal of Molecular Sciences. - 22 : 3 (2021), p. 1-23. -
További szerzők:Holzknecht, Jeanett Galgóczy László (1950-) Tóth Liliána Poór Péter Ördög Attila Váradi Györgyi Kühbacher, Alexander Borics Attila Tóth Gábor K. Marx, Florentine Batta Gyula (1953-) (molekula-szerkezet kutató)
Pályázati támogatás:NKFIH FK 134343
Egyéb
NKFIH PD 134284
Egyéb
GINOP-2.3.2-15-2016-00008
GINOP
GINOP-2.3.3-15-2016-00004
GINOP
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4.

001-es BibID:BIBFORM076180
035-os BibID:(cikkazonosító)e0204825 (WOS)000447417900013 (Scopus)85055079512
Első szerző:Fizil Ádám (biológus)
Cím:Calcium binding of the antifungal protein PAF: Structure, dynamics and function aspects by NMR and MD simulations / Ádám Fizil, Christoph Sonderegger, András Czajlik, Attila Fekete, István Komáromi, Dorottya Hajdu, Florentine Marx, Gyula Batta
Dátum:2018
ISSN:1932-6203
Megjegyzések:Calcium ions (Ca2+) play an important role in the toxicity of the cysteine-rich and cationic antifungal protein PAF from Penicillium chrysogenum: high extracellular Ca2+ levels reduce the toxicity of PAF in the sensitive model fungus Neurospora crassa in a concentration dependent way. However, little is known about the mechanistic details of the Ca2+ ion impact and the Ca2+ binding capabilities of PAF outside the fungal cell, which might be the reason for the activity loss. Using nuclear magnetic resonance (NMR), isothermal titration calorimetry and molecular dynamics (MD) simulations we demonstrated that PAF weakly, but specifically binds Ca2+ ions. MD simulations of PAF predicted one major Ca2+ binding site at the C-terminus involving Asp53 and Asp55, while Asp19 was considered as putative Ca2+ binding site. The exchange of Asp19 to serine had little impact on the Ca2+ binding, however caused the loss of antifungal activity, as was shown in our recent study. Now we replaced the C-terminal aspartates and expressed the serine variant PAF{D53S/D55S}. The specific Ca2+ binding affinity of PAF{D53S/D55S} decreased significantly if compared to PAF, whereas the antifungal activity was retained. To understand more details of Ca2+ interactions, we investigated the NMR and MD structure/dynamics of the free and Ca2+-bound PAF and PAF{D53S/D55S}. Though we found some differences between these protein variants and the Ca2+ complexes, these effects cannot explain the observed Ca2+ influence. In conclusion, PAF binds Ca2+ ions selectively at the C-terminus; however, this Ca2+ binding does not seem to play a direct role in the previously documented modulation of the antifungal activity of PAF.
Tárgyszavak:Természettudományok Kémiai tudományok idegen nyelvű folyóiratközlemény külföldi lapban
folyóiratcikk
Megjelenés:Plos One. - 13 : 10 (2018), p. 1-19. -
További szerzők:Sonderegger, Christoph Czajlik András (1975-) (gyógyszerész) Fekete Attila (1983-) (vegyész) Komáromi István (1957-) (vegyész, molekuláris biológus, biokémikus) Hajdu Dorottya (1987-) (biológus) Marx, Florentine Batta Gyula (1953-) (molekula-szerkezet kutató)
Pályázati támogatás:ANN 110821
OTKA
TÁMOP-4.2.1/B-09/1/KONV-2010-0007
TÁMOP
GINOP-2.3.2-15-2016-00008
GINOP
GINOP-2.3.3-15-2016-00004
GINOP
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5.

001-es BibID:BIBFORM058525
Első szerző:Fizil Ádám (biológus)
Cím:"Invisible" Conformers of an Antifungal Disulfide Protein Revealed by Constrained Cold and Heat Unfolding, CEST-NMR Experiments, and Molecular Dynamics Calculations / Ádám Fizil, Zoltán Gáspári, Terézia Barna, Florentine Marx, Gyula Batta
Dátum:2015
ISSN:0947-6539
Megjegyzések:Transition between conformational states in proteins is being recognized as a possible key factor of function. In support of this, hidden dynamic NMR structures were detected in several cases up to populations of a few percent. Here, we show by two- and three-state analysis of thermal unfolding, that the population of hidden states may weight 20?40?% at 298 K in a disulfide-rich protein. In addition, sensitive 15N-CEST NMR experiments identified a low populated (0.15?%) state that was in slow exchange with the folded PAF protein. Remarkably, other techniques failed to identify the rest of the NMR "dark matter". Comparison of the temperature dependence of chemical shifts from experiments and molecular dynamics calculations suggests that hidden conformers of PAF differ in the loop and terminal regions and are most similar in the evolutionary conserved core. Our observations point to the existence of a complex conformational landscape with multiple conformational states in dynamic equilibrium, with diverse exchange rates presumably responsible for the completely hidden nature of a considerable fraction.
Tárgyszavak:Természettudományok Kémiai tudományok idegen nyelvű folyóiratközlemény külföldi lapban
conformation analysis
molecular modeling
MR spectroscopy
protein folding
protein structures
Megjelenés:Chemistry-A European Journal 21 : 13 (2015), p. 5136-5144. -
További szerzők:Gáspári Zoltán Barna Teréz (1963-) (vegyész) Marx, Florentine Batta Gyula (1953-) (molekula-szerkezet kutató)
Pályázati támogatás:TÁMOP-4.2.4.A/2-11-1-2012-0001
TÁMOP
TÁMOP-4.2.1.B-11/2/KMR-2011-0002
TÁMOP
NF 104198, ANN 110821, K 105459
OTKA
NIIFI 10038
Egyéb
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6.

001-es BibID:BIBFORM127417
035-os BibID:(Scopus)85217647241 (WoS)001418685000001
Első szerző:Gai, Jiawei
Cím:Small disulfide proteins with antifungal impact: NMR experimental structures as compared to models of alphafold versions / Jiawei Gai, Márk File, Réka Erdei, András Czajlik, Florentine Marx, László Galgóczy, Györgyi Váradi, Gyula Batta
Dátum:2025
ISSN:1661-6596 1422-0067
Megjegyzések:In response to the growth of emerging resistance to conventional antifungal drugs, antifungal proteins (AFPs) of filamentous Ascomycetes origin have been discovered in recent years. Understanding the structure of AFPs is crucial for elucidating their antifungal mechanisms and developing new therapeutic agents. While nuclear magnetic resonance (NMR) has proven effective in determining the structures of small proteins, some AFP structures remain unresolved, necessitating the use of alternative prediction methods. Through bioinformatics analysis and heatmaps of amino acid sequence identity and similarity matrix, we categorized AFPs into three major classes and six subcategories, revealing structural and bioactivity differences. We employed AlphaFold (AF) to predict the 3D structures of six different AFPs, with predictions compared to NMR-derived structures. The results demonstrated a high degree of consistency between AF and NMR structures, with AF excelling in structural quality assessment and accurately capturing complex disulfide bond patterns. Both AF2 and AF3 models outperform the NMR model in overall structural quality and coherence, with AF3 showing the best performance. However, the limitations of AF should be considered, including its reduced accuracy in predicting multi-metal ion complexes, suboptimal performance in highly flexible or disordered regions, and its inability to account for multiple conformers, as it generates only a single dominant structure. Moreover, while AF3 accurately predicts all disulfide bond patterns, AF2 falls short in this regard. This study verifies the reliability of AF in the structural prediction of cysteine-rich AFPs while highlighting these constraints, offering important support for the rational design of new protein-based antifungal drugs.
Tárgyszavak:Természettudományok Kémiai tudományok idegen nyelvű folyóiratközlemény külföldi lapban
folyóiratcikk
antifungal proteins
AlphaFold
NMR
disulfide proteins
mini-protein
Megjelenés:International Journal of Molecular Sciences. - 26 : 3 (2025), p. 1-25. -
További szerzők:File Márk Erdei Réka Czajlik András (1975-) (gyógyszerész) Marx, Florentine Galgóczy László (1950-) Váradi Györgyi Batta Gyula (1953-) (molekula-szerkezet kutató)
Pályázati támogatás:NKFIH FK 134343
Egyéb
NKFIH K 146131
Egyéb
GINOP-2.3.2-15-2016-00008
GINOP
GINOP-2.3.3-15-2016-00004
GINOP
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7.

001-es BibID:BIBFORM077292
035-os BibID:(WoS)000466621200053 (Scopus)85061541042
Első szerző:Hajdu Dorottya (biológus)
Cím:Solution structure and novel insights into phylogeny and mode of action of the Neosartorya (Aspergillus) fischeri antifungal protein (NFAP) / Hajdu Dorottya, Huber Anna, Czajlik András, Tóth Liliána, Kele Zoltán, Kocsubé Sándor, Fizil Ádám, Marx Florentine, Galgóczy László, Batta Gyula
Dátum:2019
ISSN:0141-8130
Megjegyzések:Small, cysteine-rich and cationic antifungal proteins fromnatural sources are promising candidates for the development of novel treatment strategies to prevent and combat infections caused by drug-resistant fungi. However, limited information about their structure and antifungal mechanism hampers their future applications. In the present study, we determined the solution structure, dynamics and associated solvent areas of the Neosartorya (Aspergillus) fischeri antifungal protein NFAP. Genome mining within the genus revealed the presence of orthologous genes in N. fischeri and Neosartorya spathulata, and genes encoding closely related proteins can be found in Penicillium brasiliensis and Penicillium oxalicum. We show that the tertiary structure of these putative proteins can be resolved using the structure of NFAP as reliable template for in silico prediction. Localization studies with fluorescence-labelled protein pointed at an energy-dependent uptake mechanism of NFAP in the sensitive model fungus Neurospora crassa and subsequent cytoplasmic localization coincided with cell-death induction. The presented results contribute to a better understanding of the structure/function relationship of NFAP and related proteins and pave the way towards future antifungal drug development.
Tárgyszavak:Természettudományok Kémiai tudományok idegen nyelvű folyóiratközlemény külföldi lapban
folyóiratcikk
Neosartorya (Aspergillus) fischeri antifungal
protein (NFAP)
Nuclear magnetic resonance (NMR)
Antifungal mechanism
Megjelenés:International Journal of Biological Macromolecules. - 129 (2019), p. 511-522.
További szerzők:Huber Anna Czajlik András (1975-) (gyógyszerész) Tóth Liliána Kele Zoltán Kocsubé Sándor Fizil Ádám (1988-) (biológus) Marx, Florentine Galgóczy László (1950-) Batta Gyula (1953-) (molekula-szerkezet kutató)
Pályázati támogatás:GINOP-2.3.2-15-2016-00008
GINOP
GINOP-2.3.3-15-2016-00004
GINOP
PD 120808
OTKA
ANN 122833
OTKA
ANN 110821
OTKA
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8.

001-es BibID:BIBFORM018743
Első szerző:Hegedűs Nikoletta
Cím:The small molecular mass antifungal protein of Penicillium chrysogenum - a mechanism of action oriented review / Nikoletta Hegedűs, Éva Leiter, Barbara Kovács, Valéria Tomori, Nak-Jung Kwon, Tamás Emri, Florentine Marx, Gyula Batta, László Csernoch, Hubertus Haas, Jae-Hyuk Yu, István Pócsi
Dátum:2011
ISSN:0233-111X
Tárgyszavak:Természettudományok Biológiai tudományok idegen nyelvű folyóiratközlemény külföldi lapban
Molekulatudomány
Megjelenés:Journal of Basic Microbiology. - 51 : 6 (2011), p. 561-571. -
További szerzők:Leiter Éva (1976-) (biológus) Kovács Barbara Tomori Valéria Kwon, Nak-Jung Emri Tamás (1969-) (biológus) Marx, Florentine Batta Gyula (1953-) (molekula-szerkezet kutató) Csernoch László (1961-) (élettanász) Haas, Hubertus Yu Jae-Hyuk Pócsi István (1961-) (vegyész)
Pályázati támogatás:TÁMOP-4.2.1/B-09/1/KONV-2010-0007
TÁMOP
Szerkezeti biológia és molekuláris felismerés
CK77515
OTKA
NK68578
OTKA
Internet cím:Intézményi repozitóriumban (DEA) tárolt változat
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9.

001-es BibID:BIBFORM086163
035-os BibID:(WoS)000537573200009 (Scopus)85081678362
Első szerző:Huber Anna
Cím:Two small, cysteine-rich and cationic antifungal proteins from Penicillium chrysogenum : a comparative study of PAF and PAFB / A. Huber, L. Galgóczy, G. Váradi, J. Holzknecht, A. Kakar, N. Malanovic, R. Leber, J. Koch, M. A. Keller, G. Batta, G. K. Tóth, F. Marx
Dátum:2020
ISSN:0005-2736
Megjegyzések:The filamentous fungus Penicillium chrysogenum Q176 secretes the antimicrobial proteins (AMPs) PAF and PAFB, which share a compact disulfide-bond mediated, beta-fold structure rendering them highly stable. These two AMPs effectively inhibit the growth of human pathogenic fungi in micromolar concentrations and exhibit antiviral potential without causing cytotoxic effects on mammalian cells in vitro and in vivo. The antifungal mechanism of action of both AMPs is closely linked to - but not solely dependent on - the lipid composition of the fungal cell membrane and requires a strictly regulated protein uptake into the cell, indicating that PAF and PAFB are not canonical membrane active proteins. Variations in their antifungal spectrum and their killing dynamics point towards a divergent mode of action related to their physicochemical properties and surface charge distribution. In this review, we relate characteristic features of PAF and PAFB to the current knowledge about other AMPs of different sources. In addition, we present original data that have never been published before to substantiate our assumptions and provide evidences that help to explain and understand better the mechanistic function of PAF and PAFB. Finally, we underline the promising potential of PAF and PAFB as future antifungal therapeutics.
Tárgyszavak:Természettudományok Kémiai tudományok idegen nyelvű folyóiratközlemény külföldi lapban
folyóiratcikk
Antimicrobial proteins and peptides
Penicillium chrysogenum
beta-Fold structure
gamma-Core
Fungal membrane lipids
Endocytosis
Apoptosis
Megjelenés:Biochimica et Biophysica Acta (BBA). Biomembranes. - 1862 : 8 (2020), p. 1-14. -
További szerzők:Galgóczy László (1950-) Váradi Györgyi Holzknecht, Jeanett Kakar, A. Malanovic, N. Leber, R. Koch, J. Keller, M. A. Batta Gyula (1953-) (molekula-szerkezet kutató) Tóth Gábor K. Marx, Florentine
Pályázati támogatás:GINOP-2.3.2-15-2016-00008
GINOP
GINOP-2.3.3-15-2016-00004
GINOP
GINOP-2.3.2-15-2016-00014
GINOP
NKFI 20391-3/2018/FEKUSTRAT
Egyéb
TUDFO/47138-1/2019-ITM FIKP
Egyéb
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10.

001-es BibID:BIBFORM072198
035-os BibID:(Cikkazonosító)1751 (WOS)000423429800003 (Scopus)85041371349
Első szerző:Huber Anna
Cím:New Antimicrobial Potential and Structural Properties of PAFB: A Cationic, Cysteine-Rich Protein from Penicillium chrysogenum Q176 / Anna Huber, Dorottya Hajdu, Doris Bratschun-Khan, Zoltán Gáspári, Mihayl Varbanov, Stéphanie Philippot, Ádám Fizil, András Czajlik, Zoltán Kele, Christoph Sonderegger, László Galgóczy, Andrea Bodor, Florentine Marx, Gyula Batta
Dátum:2018
ISSN:2045-2322
Megjegyzések:Small, cysteine-rich and cationic proteins with antimicrobial activity are produced by diverse organisms of all kingdoms and represent promising molecules for drug development. The ancestor of all industrial penicillin producing strains, the ascomycete Penicillium chryosgenum Q176, secretes the extensively studied antifungal protein PAF. However, the genome of this strain harbours at least two more genes that code for other small, cysteine-rich and cationic proteins with potential antifungal activity. In this study, we characterized the pafB gene product that shows high similarity to PgAFP from P. chrysogenum R42C. Although abundant and timely regulated pafB gene transcripts were detected, we could not identify PAFB in the culture broth of P. chrysogenum Q176. Therefore, we applied a P. chrysogenum-based expression system to produce sufficient amounts of recombinant PAFB to address unanswered questions concerning the structure and antimicrobial function. Nuclear magnetic resonance (NMR)-based analyses revealed a compact [beta]-folded structure, comprising five [beta]-strands connected by four solvent exposed and flexible loops and an "abcabc" disulphide bond pattern. We identified PAFB as an inhibitor of growth of human pathogenic moulds and yeasts. Furthermore, we document for the first time an anti-viral activity for two members of the small, cysteine-rich and cationic protein group from ascomycetes.
Tárgyszavak:Természettudományok Kémiai tudományok idegen nyelvű folyóiratközlemény külföldi lapban
folyóiratcikk
Megjelenés:Scientific Reports. - 8 : 1 (2018), p. 1751. -
További szerzők:Hajdu Dorottya (1987-) (biológus) Bratschun-Khan, Doris Gáspári Zoltán Varbanov, Mihayl Philippot, Stéphanie Fizil Ádám (1988-) (biológus) Czajlik András (1975-) (gyógyszerész) Kele Zoltán Sonderegger, Christoph Galgóczy László (1950-) Bodor Andrea Marx, Florentine Batta Gyula (1953-) (molekula-szerkezet kutató)
Pályázati támogatás:ANN 110821
OTKA
GINOP-2.3.2-15-2016-00008
GINOP
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Intézményi repozitóriumban (DEA) tárolt változat
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11.

001-es BibID:BIBFORM066900
035-os BibID:(WoS)000387640700002 (Scopus)84995519563
Első szerző:Sonderegger, Christoph
Cím:A Penicillium chrysogenum-based expression system for the production of small, cysteine-rich antifungal proteins for structural and functional analyses / Christoph Sonderegger, László Galgóczy, Sandra Garrigues, Ádám Fizil, Attila Borics, Paloma Manzanares, Nikoletta Hegedüs, Anna Huber, Jose F. Marcos, Gyula Batta, Florentine Marx
Dátum:2016
ISSN:1475-2859
Megjegyzések:Small, cysteine-rich and cationic antifungal proteins (APs) from filamentous ascomycetes, such as NFAP from Neosartorya fischeri and PAF from Penicillium chrysogenum, are promising candidates for novel drug development. A prerequisite for their application is a detailed knowledge about their structure-function relation and mode of action, which would allow protein modelling to enhance their toxicity and specificity. Technologies for structure analyses, such as electronic circular dichroism (ECD) or NMR spectroscopy, require highly purified samples and in case of NMR milligrams of uniformly 15N-/13C-isotope labelled protein. To meet these requirements, we developed a P. chrysogenum-based expression system that ensures sufficient amount and optimal purity of APs for structural and functional analyses.RESULTS:The APs PAF, PAF mutants and NFAP were expressed in a P. chrysogenum ?paf mutant strain that served as perfect microbial expression factory. This strain lacks the paf-gene coding for the endogenous antifungal PAF and is resistant towards several APs from other ascomycetes. The expression of the recombinant proteins was under the regulation of the strong paf promoter, and the presence of a paf-specific pre-pro sequence warranted the secretion of processed proteins into the supernatant. The use of defined minimal medium allowed a single-step purification of the recombinant proteins. The expression system could be extended to express PAF in the related fungus Penicillium digitatum, which does not produce detectable amounts of APs, demonstrating the versatility of the approach. The molecular masses, folded structures and antifungal activity of the recombinant proteins were analysed by ESI-MS, ECD and NMR spectroscopy and growth inhibition assays.CONCLUSION:This study demonstrates the implementation of a paf promoter driven expression cassettes for the production of cysteine-rich, cationic, APs in different Penicillium species. The system is a perfect tool for the generation of correctly folded proteins with high quality for structure-function analyses.KEYWORDS:Antifungal proteins; Electronic circular dichroism (ECD) spectroscopy; NFAP; Neosartorya fischeri; Nuclear magnetic resonance (NMR); PAF; Penicillium chrysogenum; Penicillium digitatum; Recombinant protein production
Tárgyszavak:Természettudományok Biológiai tudományok idegen nyelvű folyóiratközlemény külföldi lapban
Megjelenés:Microbial Cell Factories. - 15 : 192 (2016), p. 1-14. -
További szerzők:Galgóczy László (1950-) Garrigues, Sandra Fizil Ádám (1988-) (biológus) Borics Attila Manzanares, Paloma Hegedűs Nikoletta Huber Anna Marcos, Jose F. Batta Gyula (1953-) (molekula-szerkezet kutató) Marx, Florentine
Pályázati támogatás:ANN110821
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Intézményi repozitóriumban (DEA) tárolt változat
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12.

001-es BibID:BIBFORM066897
035-os BibID:(cikkazonosító)e0169920 (WOS)000391844200058 (Scopus)85009084271
Első szerző:Sonderegger, Christoph
Cím:D19S Mutation of the Cationic, Cysteine-Rich Protein PAF: Novel Insights into Its Structural Dynamics, Thermal Unfolding and Antifungal Function / Christoph Sonderegger, Ádám Fizil, Laura Burtscher, Dorottya Hajdu, Alberto Muñoz, Zoltán Gaáspári, Nick D. Read, Gyula Batta, Florentine Marx
Dátum:2017
ISSN:1932-6203
Megjegyzések:The cysteine-rich, cationic, antifungal protein PAF is abundantly secreted into the culture supernatant of the filamentous Ascomycete Penicillium chrysogenum. The five ?-strands of PAF form a compact ?-barrel that is stabilized by three disulphide bonds. The folding of PAF allows the formation of four surface-exposed loops and distinct charged motifs on the protein surface that might regulate the interaction of PAF with the sensitive target fungus. The growth inhibitory activity of this highly stable protein against opportunistic fungal pathogens provides great potential in antifungal drug research. To understand its mode of action, we started to investigate the surface-exposed loops of PAF and replaced one aspartic acid at position 19 in loop 2 that is potentially involved in PAF active or binding site, with a serine (Asp19 to Ser19). We analysed the overall effects, such as unfolding, electrostatic changes, sporadic conformers and antifungal activity when substituting this specific amino acid to the fairly indifferent amino acid serine. Structural analyses revealed that the overall 3D solution structure is virtually identical with that of PAF. However, PAFD19S showed slightly increased dynamics and significant differences in the surface charge distribution. Thermal unfolding identified PAFD19S to be rather a two-state folder in contrast to the three-state folder PAF. Functional comparison of PAFD19S and PAF revealed that the exchange at residue 19 caused a dramatic loss of antifungal activity: the binding and internalization of PAFD19S by target cells was reduced and the protein failed to trigger an intracellular Ca2+ response, all of which are closely linked to the antifungal toxicity of PAF. We conclude that the negatively charged residue Asp19 in loop 2 is essential for full function of the cationic protein PAF.
Tárgyszavak:Természettudományok Kémiai tudományok idegen nyelvű folyóiratközlemény külföldi lapban
folyóiratcikk
Megjelenés:Plos One. - 12 : 1 (2017), p. e0169920-1 - e0169920-21. -
További szerzők:Fizil Ádám (1988-) (biológus) Burtscher, Laura Hajdu Dorottya (1987-) (biológus) Munoz, Alberto Gaáspári Zoltán Read, Nick D. Batta Gyula (1953-) (molekula-szerkezet kutató) Marx, Florentine
Pályázati támogatás:ANN110821
OTKA
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Intézményi repozitóriumban (DEA) tárolt változat
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