Antifungal protein J

Background: Alpha-1-syntrophin (SNTA1) is emerging as a novel modulator of the actin cytoskeleton. SNTA1 binds to F-actin and regulates intracellular localization and activity of various actin organizing signaling molecules. Aberration in syntrophin signaling has been closely linked with deregulated growth connected to tumor development/metastasis and its abnormal over expression has been observed in breast cancer. In the present work the effect of jasplakinolide, an actin-binding cyclodepsipeptide, on the SNTA1 protein activity and SNTA1 mediated downstream cellular events was studied in MDA-MB-231 breast cancer cell line. Methods: SNTA1 protein levels and phosphorylation status were determined in MDA-MB-231 cells post jasplakinolide exposure using western blotting and immunoprecipitation techniques respectively. MDA-MB-231 cells were transfected with WT SNTA1 and DM SNTA1 (Y215/229 phospho mutant) and simultaneously treated with jasplakinolide. The effect of jasplakinolide and SNTA1 protein on cell migration was determined using the boyden chamber assay. Results: Jasplakinolide treatment decreases proliferation of MDA-MB-231 cells in both dose and time dependent manner. Results suggest that subtoxic doses of jasplakinolide induce morphological changes in MDA-MB-231 cells from flat spindle shape adherent cells to round weakly adherent forms. Mechanistically, jasplakinolide treatment was found to decrease SNTA1 protein levels and its tyrosine phosphorylation status. Moreover, migratory potential of jasplakinolide treated cells was significantly inhibited in comparison to control cells. Conclusion: Our results demonstrate that jasplakinolide inhibits cell migration by impairing SNTA1 functioning in breast cancer cells.

Biological control to prevent fungal plant diseases offers an alternative approach to facilitate sustainable agriculture. Since the chitin in fungal cell walls is a target for biocontrol agents, chitinases are one of the important antifungal molecules. In this study, the aim was to investigate a new chitinase isolated from a fluvial soil bacterium and to show the antifungal activity of the characterized chitinase by comparing the three common methods. The bacterium with the highest chitinase activity was identified as Aeromonas sp. by 16 S rRNA sequence analysis. Following the determination of the optimum enzyme production time, the enzyme was partially purified, and the physicochemical parameters of the enzyme were investigated. In the antifungal studies, direct Aeromonas sp. BHC02 cells or partially purified chitinase were used. As a result, in the first method in which the Aeromonas sp. BHC02 cells were spread on the surface of petri dishes, no zone formation was observed around the test fungi spotted on the surface. However, zone formation was observed in the methods in which the antifungal activity was investigated using the partially purified chitinase enzyme. For example, in the second method, the enzyme was spread on the surface of PDA, and zone formation was observed only around Penicillum species among the test fungi spotted on the surface. In the third method, in which the necessary time was given for the formation of mycelium of the test fungi, it was observed that the growth of Fusarium solani, Alternaria alternata and Botrytis cinerea was inhibited by the partially purified chitinase. This study concludes that the results of the antifungal activities depend on the method used and all fungal chitins cannot be degraded with one strain's chitinase. Depending on the variety of chitin, some fungi can be more resistant.

Antifungal proteins and peptides have drawn the attention of numerous plant biologists and Clinicians, owing to their potential value in protecting commercial crops as well as preventing fungal infections in humans. Various proteins and peptides, such as glucanases, chitinases, chitinase-like proteins, lectins, peroxidases, defensins, and lipid transfer proteins have antifungal activities. Thaumatin is a protein from a West African plant Thaumatococcus danielli that is sweet in taste but does not exhibit antifungal activities. Despite the structural similarities between thaumatins and thaumatin-like proteins (TLPs), TLPs are not sweet in taste, unlike thaumatins. We developed a thaumatin-like protein database of various organisms. TLPs are pathogenesis-related proteins (PR) with molecular masses of 20-26 kDa. The amino acid residues of TLPs involved in an antifungal activity remain obscure and make it hard to receive comprehensive information on TLPs. The biggest problem in the wine industry is white haze, an undesirable feature of high-quality wine. Hence, the problem may be figured out with the easy accessibility of amino acid sequences and to generate infest resistant crops. Overall, we aimed to produce a freely accessible TLP database ( https://tlpdb.cftri.com ) that would provide substantive information in understanding the mechanistic facet of TLPs. Briefly, TLPdb contains sequences, structures, and amino acid compositions of validated, published TLP protein sequences (from the plant, fungal as well as animal sources). Thus, this work may yield valuable information that may be useful in understanding the mechanistic aspects of TLP activity and in the evolution of antifungal proteins and fungal resistant crops. TLPdb is a comprehensive thaumatin-like protein resource database of various organisms. The database can serve as a unique Bioinformatics tool for understanding the TLPs. This further may help in understanding and the development of fungal resistant crops. TLPdb is freely available at https://tlpdb.cftri.com .