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  • Title: First simultaneous isolation of a ribosome inactivating protein and an antifungal protein from a mushroom (Lyophyllum shimeji) together with evidence for synergism of their antifungal effects.
    Author: Lam SK, Ng TB.
    Journal: Arch Biochem Biophys; 2001 Sep 15; 393(2):271-80. PubMed ID: 11556814.
    Abstract:
    From the fruiting bodies of the mushroom Lyophyllum shimeji, a novel ribosome inactivating protein with a molecular weight of 20 kDa and exhibiting antifungal activity against Physalospora piricola (IC(50) = 2.5 microM) and Coprinus comatus was isolated. The protein, designated lyophyllin, was purified by ion exchange chromatography on CM-cellulose, affinity chromatography on Affi-gel Blue Gel, and then ion exchange chromatography on Mono S. Lyophyllin possessed an N-terminal sequence with some similarity to those of plant ribosome-inactivating proteins. It inhibited translation in rabbit reticulocyte lysate with an IC(50) of 1 nM, thymidine uptake by murine splenocytes with an IC(50) of 1 microM and HIV-1 reverse transcriptase activity with an IC(50) of 7.9 nM. Lyophyllin did not manifest ribonuclease or hemagglutinating activity. An antifungal protein, designated Lyophyllum antifungal protein (LAP), with a molecular weight of 14 kDa, and an N-terminal sequence somewhat analogous to those of angiosperm thaumatin-like proteins and thaumatins and an inactive variant of the ubiquitin-conjugating enzyme, was first isolated from Lyophyllum shimeji. LAP was adsorbed on CM-cellulose, Affi-gel blue gel, and Mono S. LAP exerted antifungal activity against P. piricola (IC(50) = 70 nM) and Mycosphaerella arachidicola but not against Rhizoctonia solani, Colletotrichum gossypii, and Coprinus comatus. It exerted very low translation inhibitory activity in a rabbit reticulocyte lysate system (IC(50) = 70 microM) and negligible ribonuclease activity toward yeast transfer RNA and hemagglutinating activity toward rabbit erythrocytes. It inhibited HIV-1 reverse transcriptase with an IC(50) of about 5.2 nM. A synergism in antifungal activities of LAP and lyophyllin against P. piricola was demonstrable.
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