Proteins associated with the cell envelope of Trichoderma reesei: A proteomic approach

Abstract

A total of 220 cell envelope-associated proteins were successfully extracted and separated from Trichoderma reesei mycelia actively synthesizing and secreting proteins and from mycelia in which the secretion of proteins are low. Altogether 56 spots were examined by nanoelectrospray tandem mass spectrometry and amino acid sequence was obtained for 32 spots. From these, 20 spots were identified by Advanced BLAST searches against all databases available to BLAST. The most abundant protein in both types of mycelia was HEX1, the major protein in Woronin body, a structure unique to filamentous fungi. Other proteins identified were vacuolar protease A, enolase, glyceraldehyde-3-phosphate dehydrogenase, transaldolase, protein disulfide isomerase, mitochondrial outer membrane porin, diphosphate kinase and translation elongation factor beta. Partial short amino acid sequence obtained from some proteins did not allow them to be assigned to a specific protein in the database by BLAST search. In some cases, the tandem mass spectrometry spectra were too complicated to be able to assign an amino acid sequence with certainty. The number of spots (12) giving a clear signal but finding no match in the databases suggests that a majority of proteins associated with a filamentous fungal cell wall, are novel. Some technical problems related to protein isolation are also discussed.

Proteomic response of Trichoderma aggressivum f. europaeum to Agaricus bisporus tissue and mushroom compost

“…Sequence analysis of Hex 1 from reveals that it has a peroxisome targeting signal and a high level of conservation between fungal species (Curach et al 2004). In other Trichoderma species it has been shown to account for a significant amount of the total protein associated with the cell envelope (Lim et al 2001) and is differentially produced in response to different carbon sources (Curach et al 2004) and under biological control conditions (Marra et al 2006) but no change was observed in Hex 1 abundance in either experimental treatment in this study.…”

Pathogen to Endophytic Transmission in Fungi- A Proteomics Approach

“…Some of the fungal species are of crucial significance for their physiological and symbiotic abilities like mycorrhizal associations which at times are crucial for plant growth, development and fruiting [6]. Proteomic studies of filamentous fungi were started by Lim, et al on Trichoderma reesei cell envelope proteins and on Aspergillus fumigates glycosyl phosphatidyl inositol-anchored proteins by Bruneau, et al [7,8].…”

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

“…Thus, the use of liquid nitrogen to decrease warming of the disruption systems is a widely used method. The main cell-breaking system is the traditional pre-chilled mortar grinding due to its efficiency against the fungal cell wall (Cobos et al, 2010;Lu et al, 2010;Vödisch et al, 2011), although waring blender machines (Lim et al, 2001), or glass bead beating systems, either combined with a 10 mM Tris-HCl buffer (Oh et al, 2010) or with a phenol buffer (Coumans et al, 2010;Vodisch et al, 2011), have been successfully applied. After the breaking step, the protein solubilisation buffer always includes a protease inhibitor [e.g.…”

“…Thus, Trichoderma genus has generated a special attention as active agent for biological control of plant pathogenic fungi. Proteins associated with the cell envelope of Trichoderma reesei were analysed by 2D electrophoresis allowing the location of 220 proteins and the identification of 32 spots by nanoelectrospray tandem mass spectrometry [Q-TOF MS (Micromass)] and amino acid sequence (Lim et al, 2001). The most abundant protein was HEX1, the major protein in Woronin body, suggesting that this structure unique to filamentous fungi is linked to the cell envelope.…”

“…Cell envelope 2DE / ESI-Q-TOF Biocontrol agent Lim et al, 2001 Table 3. Summary of the major proteomes identified from filamentous fungi using different proteomic approaches.…”