Iron-regulated proteins inPhanerochaete chrysosporium andLentinula edodes: Differential analysis by sodium dodecyl sulfate polyacrylamide gel electrophoresis and two-dimensional polyacrylamide gel electrophoresis profiles

Abstract

Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and two-dimensional gel electrophoresis (2-DE) were used to identify iron-responsive proteins in the white-rot species (Phanerochaete chrysosporium and Lentinula edodes), by comparing the differential patterns of cellular and membrane proteins obtained from iron-sufficient and iron-deficient mycelia. Six cellular proteins induced by iron restriction have been observed in SDS-PAGE for P. chrysosporium and twelve for L. edodes. In 2-DE, the numbers of iron-restricted induced proteins were 12 and 9, respectively, in a resolution range of 15-60 kDa and pI 4.5-8.1. SDS-PAGE for the plasma membrane protein did not show differences, whereas the outer-membrane protein profile showed 6 and 5 proteins induced by iron depletion in P. chrysosporium and L. edodes, respectively. The results presented here are important data to unravel mechanisms of biosynthesis and/or transport of the iron-complexing agents in ligninolytic fungi and to further correlate them to the ligninolytic processes.

Proteome analysis of Aspergillus ochraceus

“…Several researchers (Melin et al 2002;Nandakumar et al 2003;Grinyer et al 2004;Strom et al 2005) used mechanical lysis via glass beads to liberate cytoplasmic proteins, and this approach has been reported to be more efficient than either chemical or enzymatic extraction methods (Nandakumar and Marten 2002). However, the most widely used method for extraction seems to be grinding in liquid nitrogen using mortar and pestle (Hernandez-Macedo et al 2002;Grinyer et al 2005;Shimizu and Wariishi 2005;Kniemeyer et al 2006;Yajima and Kav 2006).…”

Pathogen to Endophytic Transmission in Fungi- A Proteomics Approach

“…Hernández-Macedo, et al carried out fungal protein vizualisation by 2DPAGE [41]. A filamentous fungi used as a model organism is Aspergillus sp.…”

Proteomics Methodology Applied to the Analysis of Filamentous Fungi - New Trends for an Impressive Diverse Group of Organisms

“…Now, the major challenge in modern fungal biology is to understand the expression, function and regulation of the entire set of proteins encoded by fungal genomes. The combination of 2D-PAGE and mass spectrometry for the identification of proteins from filamentous fungi was first used by Grinyer and co-workers (2004), although reports on 2D-PAGE visualization of fungal proteins had been previously carried out (Hernández-Macedo et al, 2002). Combination of 2D-PAGE with both MALDI-TOF (Kratos Analytical AXIMA CFR plus) and LC-MS/MS [LCQ Deca IT MS (Thermo Finnigan)] techniques, allowed the initial identification of 25 proteins (out of 96 attempted) from the hundreds of proteins resolved in the initial proteome map of Trichoderma harzianum, which was performed using whole-cell protein extracts.…”

“…For example, the discovery of novel sources of host resistance, the evolution of strains due to the industrial improvement or the effects of carbon sources, antifungal drugs and gene deletion have been analysed in detail (Fernández-Acero et al, 2010;Jami et al, 2010a;Cagas et al, 2011b). One of the earliest intracellular filamentous fungal proteomic studies was performed on the wood-degrading fungi Phanerochaete chrysosporium and Lentinula edodes (Hernández-Macedo et al, 2002). These authors used 2D-PAGE to compare cytoplasmic protein expression patterns in the presence or absence of iron and although they visualized 21 proteins related to iron uptake, the identification of such proteins was deficient.…”