Direct identification of hydrophobins and their processing in Trichoderma using intact-cell MALDI-TOF MS.
Intact-cell MS (ICMS) was applied for the direct detection of hydrophobins in various species and strains of Hypocrea/Trichoderma. In both mycelia and spores, dominating peaks were identified as hydrophobins by detecting mass shifts of 8 Da of reduced and unreduced forms, the analysis of knockout mutants, and comparison with protein databases. Strain-specific processing was observed in the case of Hypocrea jecorina (anamorph Trichoderma reesei). An analysis of 32 strains comprising 29 different species of Trichoderma and Hypocrea showed hydrophobin patterns that were specific at both at the species and isolate (subspecies) levels. The method therefore permits rapid and direct detection of hydrophobin class II compositions and may also provide a means to identify Trichoderma (and other fungal) species and strains from microgram amounts of biomass without prior cultivation.
Neuhof
Torsten
T
TU Berlin, Institut für Chemie, FG Biochemie und Molekulare Biologie, Berlin, Germany.
Dieckmann
Ralf
R
Druzhinina
Irina S
IS
Kubicek
Christian P
CP
Nakari-Setälä
Tiina
T
Penttilä
Merja
M
von Döhren
Hans
H
D000595:Amino Acid Sequence; D020224:Expressed Sequence Tags; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D025301:Hyphae / Q000235:genetics / Q000378:metabolism; D035901:Hypocrea / Q000145:classification / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D009154:Mutation; D011499:Protein Processing, Post-Translational; D016415:Sequence Alignment; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D013172:Spores, Fungal / Q000737:chemistry / Q000378:metabolism; D014242:Trichoderma / Q000145:classification / Q000235:genetics / Q000378:metabolism*
D005656:Fungal Proteins