Identification of protein complexes from filamentous fungi with tandem affinity purification.
Fungal molecular biology has benefited from the enormous advances in understanding protein-protein interactions in prokaryotic or eukaryotic organisms of the past decade. Tandem affinity purification (TAP) allows the enrichment of native protein complexes from cell extracts under mild conditions. We codon-optimized tags and established TAP, previously not applicable to filamentous fungi, for the model organism Aspergillus nidulans. We could identify by this method the trimeric Velvet complex VelB/VeA/LaeA or the eight subunit COP9 signalosome. Here, we describe an optimized protocol for A. nidulans which can also be adapted to other filamentous fungi.
Abteilung Molekulare Mikrobiologie und Genetik, DFG Research Center for Molecular Physiology of the Brain (CMPB), Georg-August-Universität Göttingen, Göttingen, Germany.
Ozgür
O
Bayram
Ozlem Sarikaya
OS
Bayram
Oliver
O
Valerius
Bastian
B
Jöhnk
Gerhard H
GH
Braus
D001233:Aspergillus nidulans / Q000737:chemistry*; D001557:Benzenesulfonates / Q000737:chemistry; D002846:Chromatography, Affinity / Q000379:methods*; D002851:Chromatography, High Pressure Liquid; D030562:Databases, Protein; D004591:Electrophoresis, Polyacrylamide Gel; D005656:Fungal Proteins / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D013058:Mass Spectrometry; D059748:Proteolysis; D016622:Silver Staining; D014357:Trypsin / Q000378:metabolism
D001557:Benzenesulfonates; D005656:Fungal Proteins; D014357:Trypsin; C006796:brilliant blue