title: Linear Discriminant Analysis Identifies Mitochondrially Localized Proteins in Neurospora crassa.
abstract: Besides their role as powerhouses, mitochondria play a pivotal role in the spatial organization of numerous enzymatic functions. They are connected to the ER, and many pathways are organized through the mitochondrial membranes. Thus, the precise definition of mitochondrial proteomes remains a challenging task. Here, we have established a proteomic strategy to accurately determine the mitochondrial localization of proteins from the fungal model organism Neurospora crassa. This strategy relies on both highly pure mitochondria as well as the quantitative monitoring of mitochondrial components along their consecutive enrichment. Pure intact mitochondria were obtained by a multistep approach combining differential and density Percoll (ultra) centrifugations. When compared with three other intermediate enrichment stages, peptide sequencing and quantitative profiling of pure mitochondrial fractions revealed prototypic regulatory profiles of per se mitochondrial components. These regulatory profiles constitute a distinct cluster defining the mitochondrial compartment and support linear discriminant analyses, which rationalized the annotation process. In total, this approach experimentally validated the mitochondrial localization of 512 proteins including 57 proteins that had not been reported for N. crassa before.
authors.affiliation: Cellular Proteomics Research Group, §Central Facility for Microscopy, Helmholtz Centre for Infection Research , 38124 Braunschweig, Germany.
authors.forename: Lisette
authors.initials: L
authors.lastname: Wirsing
authors.affiliation: Cellular Proteomics Research Group, §Central Facility for Microscopy, Helmholtz Centre for Infection Research , 38124 Braunschweig, Germany.|Department of Computer Science, Ostfalia University of Applied Sciences , 38302 Wolfenbüttel, Germany.
authors.forename: Frank
authors.initials: F
authors.lastname: Klawonn
authors.forename: Wiebke Anna
authors.initials: WA
authors.lastname: Sassen
authors.forename: Heinrich
authors.initials: H
authors.lastname: Lünsdorf
authors.forename: Corinna
authors.initials: C
authors.lastname: Probst
authors.forename: Michael
authors.initials: M
authors.lastname: Hust
authors.forename: Ralf R
authors.initials: RR
authors.lastname: Mendel
authors.forename: Tobias
authors.initials: T
authors.lastname: Kruse
authors.affiliation: Cellular Proteomics Research Group, §Central Facility for Microscopy, Helmholtz Centre for Infection Research , 38124 Braunschweig, Germany.
authors.forename: Lothar
authors.initials: L
authors.lastname: Jänsch
mesh_terms: D016002:Discriminant Analysis; D005656:Fungal Proteins / Q000032:analysis* / Q000737:chemistry; D024101:Mitochondrial Proteins / Q000032:analysis* / Q000737:chemistry; D009492:Neurospora crassa / Q000737:chemistry* / Q000166:cytology*; D040901:Proteomics / Q000379:methods*
chemical_list: D005656:Fungal Proteins; D024101:Mitochondrial Proteins
keywords: Mitochondrial proteomics; Neurospora crassa; d-arabinitol dehydrogenase; heme biosynthesis; linear discriminant analysis (LDA)