TY - JOUR
T1 - Proteins associated with the cell envelope of Trichoderma reesei
T2 - a proteomic approach
AU - Lim, Dongbin
AU - Hains, Peter
AU - Walsh, Brad
AU - Bergquist, Peter
AU - Nevalainen, Helena
PY - 2001
Y1 - 2001
N2 - 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.
AB - 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.
KW - cell envelope
KW - Trichoderma reesei
KW - two-dimensional electrophoresis
KW - proteomic display
UR - http://www.scopus.com/inward/record.url?scp=0035406437&partnerID=8YFLogxK
U2 - 10.1002/1615-9861(200107)1:7<899::AID-PROT899>3.0.CO;2-#
DO - 10.1002/1615-9861(200107)1:7<899::AID-PROT899>3.0.CO;2-#
M3 - Article
C2 - 11503214
AN - SCOPUS:0035406437
SN - 1615-9853
VL - 1
SP - 899
EP - 910
JO - Proteomics
JF - Proteomics
IS - 7
ER -