Phanerochaete chrysosporium soluble proteome as a prelude for the analysis of heavy metal stress response

Abstract

A 2-D reference map in pI range 3-10 was constructed for the soluble protein fraction of Phanerochaete chrysosporium growing vegetatively under standard conditions. Functional annotation could be made for 517 spots out of 720 that were subjected to MALDI-TOF-MS analysis, according to the specific accession numbers from the P. chrysosporium genomic database. Further analysis of the data revealed 314 distinct ORFs, 118 of which yielded multiple spots on the master gel. Functional classification of the proteins was made according to the eukaryote orthologous groups defined in the organism's genome website. The functional class of PTMs, protein turnover and chaperones was represented with the highest number (63) of the identified ORFs. Six proteins were assigned to the hypothetical proteins and 29 were predicted to have a signal peptide sequence. Subcellular localization predictions were also made for the identified proteins. Of the protein spots detected on the master gel, 380 were found to be probably phosphorylated and 96 of these matched to the identified proteins. The reference map was efficiently used in the identification of the proteins differentially expressed under cadmium and copper stress. Three new ribosomal proteins as well as zinc-containing alcohol dehydrogenase, glucose-6-phosphate isomerase, flavonol/cinnamoyl-CoA reductase, H+-transporting two-sector ATPase, ribosomal protein S7, ribosomal protein S21e, elongation factor EF-1 alpha subunit were demonstrated as the most strongly induced.

Transcriptomic Responses of Phanerochaete chrysosporium to Oak Acetonic Extracts: Focus on a New Glutathione Transferase

“…This enzyme participates in the lignin biosynthesis pathway (36). Moreover, a 9-fold-elevated level of cinnamoyl-CoA reductase protein in P. chrysosporium under Cu stress suggests a role of this enzyme in stress response (37). In addition to the genes reported in Table 1, 73 genes exhibiting no homology in the databases are upregulated more than 10-fold.…”

Differential Expression in Phanerochaete chrysosporium of Membrane-Associated Proteins Relevant to Lignin Degradation

“…Now that the genome of Phanerochaete chrysosporium has been sequenced and annotated (20, 34), proteomic work has begun on this intensively researched white rot fungus. Recent studies have shown variation in the expression patterns of many extracellular and cytoplasmic enzymes relevant to ligninolysis (1,21,26,28,30,35) but have so far revealed little about the regulation of membrane-associated components. In part, this lack of information likely reflects the poor behavior of some membrane proteins during the electrophoretic separations that are generally employed (27).…”

Extracellular proteomic analysis for degradation of PAHs in source of drinking water with fusant strains

“…The supernatant was concentrated 100-fold by an Amicon 10 kDa filter at 4°C. Then TCA-acetone method was used to extract the total extracellular proteins (Ozcan et al 2007). A volume of 5 ml 10% TCA in acetone containing 0.07% b-mercaptoethanol was added and vortexed, then incubated at -20°C for 45 min and centrifuged at 15,0009g for 15 min.…”