Hydrated conidia of Metarhizium anisopliae release a family of metalloproteases.
Metarhizium anisopliae spores release isoforms of metalloprotease during hydration over a 4-day incubation period. The isoforms were identified and characterized by using one-dimensional native PAGE (1-DE nPAGE) and one-dimensional SDS non-dissociating (1-DE nSDS-PAGE) zymography. The ability of these isozymes to degrade gelatin varied as revealed by 2-D spot densitometry. 1-DE nPAGE zymography revealed five isoforms of gelatinase from Tween wash of conidia. Where as, one to three activities with different intensities appeared on gel from washing of conidia to incubation in water till day 4. The relative migrations of these activities on 1-DE nPAGE zymograms appeared as fast, medium and slow on gel. The 2-D spot densitometry of zymograms indicated isoforms have different proteolytic activity as quantified by pixel intensities. SDS-PAGE zymography indicated the release of two isozymes of Mr 103 and 12 kDa during Tween treatment of conidia. However, during the first washing step with water and incubation of spores at day 2 and 3, respectively, only 12 kDa protein was evident. Majority of these proteases were inhibited by EDTA, but stimulated by CaCl(2), and MgCl(2). The presence of isozymes in conidia and their release during hydration must have functional significance for fungi and in this case it should provide advantages to M. anisopliae in its saprobic or pathogenic modalities. To our knowledge this is the first report describing release of metalloprotease isozymes from conidia.
Qazi
Sohail S
SS
Microbial Biotechnology/Molecular Microbiology Laboratories, Department of Applied Microbiology and Food Science, University of Saskatchewan, Saskatoon, Canada. soq823@mail.usask.ca
Khachatourians
George G
GG
D000818:Animals; D018093:Gelatinases / Q000378:metabolism; D007527:Isoenzymes / Q000378:metabolism; D045726:Metalloproteases / Q000378:metabolism*; D052981:Metarhizium / Q000201:enzymology* / Q000472:pathogenicity; D013172:Spores, Fungal / Q000201:enzymology*; D014867:Water
D007527:Isoenzymes; D014867:Water; D045726:Metalloproteases; D018093:Gelatinases