title: Mitochondrial Protein Interaction Mapping Identifies Regulators of Respiratory Chain Function.
abstract: Mitochondria are essential for numerous cellular processes, yet hundreds of their proteins lack robust functional annotation. To reveal functions for these proteins (termed MXPs), we assessed condition-specific protein-protein interactions for 50 select MXPs using affinity enrichment mass spectrometry. Our data connect MXPs to diverse mitochondrial processes, including multiple aspects of respiratory chain function. Building upon these observations, we validated C17orf89 as a complex I (CI) assembly factor. Disruption of C17orf89 markedly reduced CI activity, and its depletion is found in an unresolved case of CI deficiency. We likewise discovered that LYRM5 interacts with and deflavinates the electron-transferring flavoprotein that shuttles electrons to coenzyme Q (CoQ). Finally, we identified a dynamic human CoQ biosynthetic complex involving multiple MXPs whose topology we map using purified components. Collectively, our data lend mechanistic insight into respiratory chain-related activities and prioritize hundreds of additional interactions for further exploration of mitochondrial protein function.
authors: Morgridge Institute for Research, Madison, WI 53715, USA; Department of Biochemistry, University of Wisconsin-Madison, Madison, WI 53706, USA.
authors: Brendan J
authors: BJ
authors: Floyd
authors: Department of Chemistry, University of Wisconsin-Madison, Madison, WI 53706, USA.
authors: Emily M
authors: EM
authors: Wilkerson
authors: Morgridge Institute for Research, Madison, WI 53715, USA; Department of Biochemistry, University of Wisconsin-Madison, Madison, WI 53706, USA.
authors: Mike T
authors: MT
authors: Veling
authors: Department of Chemistry, University of Wisconsin-Madison, Madison, WI 53706, USA.
authors: Catie E
authors: CE
authors: Minogue
authors: Department of Biochemistry, Medical College of Wisconsin, Milwaukee, WI 53226, USA.
authors: Chuanwu
authors: C
authors: Xia
authors: Department of Biochemistry, University of Wisconsin-Madison, Madison, WI 53706, USA.
authors: Emily T
authors: ET
authors: Beebe
authors: Department of Biochemistry, University of Wisconsin-Madison, Madison, WI 53706, USA.
authors: Russell L
authors: RL
authors: Wrobel
authors: Morgridge Institute for Research, Madison, WI 53715, USA; Department of Biochemistry, University of Wisconsin-Madison, Madison, WI 53706, USA.
authors: Holly
authors: H
authors: Cho
authors: Institute of Human Genetics, Technische Universität München, 81675 München, Germany; Institute of Human Genetics, Helmholtz Zentrum München, 85764 Neuherberg, Germany.
authors: Laura S
authors: LS
authors: Kremer
authors: Wellcome Trust Centre for Mitochondrial Research, Institute of Neuroscience, The Medical School, Newcastle University, Newcastle upon Tyne NE2 4HH, UK.
authors: Charlotte L
authors: CL
authors: Alston
authors: Department of Biochemistry, University of Wisconsin-Madison, Madison, WI 53706, USA.
authors: Katarzyna A
authors: KA
authors: Gromek
authors: Department of Biochemistry, University of Wisconsin-Madison, Madison, WI 53706, USA.
authors: Brendan K
authors: BK
authors: Dolan
authors: Department of Chemistry, University of Wisconsin-Madison, Madison, WI 53706, USA.
authors: Arne
authors: A
authors: Ulbrich
authors: Morgridge Institute for Research, Madison, WI 53715, USA; Department of Biochemistry, University of Wisconsin-Madison, Madison, WI 53706, USA.
authors: Jonathan A
authors: JA
authors: Stefely
authors: Morgridge Institute for Research, Madison, WI 53715, USA; Department of Biochemistry, University of Wisconsin-Madison, Madison, WI 53706, USA.
authors: Sarah L
authors: SL
authors: Bohl
authors: Department of Biochemistry, University of Wisconsin-Madison, Madison, WI 53706, USA.
authors: Kelly M
authors: KM
authors: Werner
authors: Morgridge Institute for Research, Madison, WI 53715, USA.
authors: Adam
authors: A
authors: Jochem
authors: Genome Center of Wisconsin, University of Wisconsin-Madison, Madison, WI 53706, USA.
authors: Michael S
authors: MS
authors: Westphall
authors: Morgridge Institute for Research, Madison, WI 53715, USA.
authors: Jarred W
authors: JW
authors: Rensvold
authors: Wellcome Trust Centre for Mitochondrial Research, Institute of Neuroscience, The Medical School, Newcastle University, Newcastle upon Tyne NE2 4HH, UK.
authors: Robert W
authors: RW
authors: Taylor
authors: Institute of Human Genetics, Technische Universität München, 81675 München, Germany; Institute of Human Genetics, Helmholtz Zentrum München, 85764 Neuherberg, Germany.
authors: Holger
authors: H
authors: Prokisch
authors: Department of Biochemistry, Medical College of Wisconsin, Milwaukee, WI 53226, USA.
authors: Jung-Ja P
authors: JP
authors: Kim
authors: Department of Chemistry, University of Wisconsin-Madison, Madison, WI 53706, USA; Department of Biomolecular Chemistry, University of Wisconsin-Madison, Madison, WI 53706, USA; Genome Center of Wisconsin, University of Wisconsin-Madison, Madison, WI 53706, USA.
authors: Joshua J
authors: JJ
authors: Coon
authors: Morgridge Institute for Research, Madison, WI 53715, USA; Department of Biochemistry, University of Wisconsin-Madison, Madison, WI 53706, USA. Electronic address: dpagliarini@morgridge.org.
authors: David J
authors: DJ
authors: Pagliarini
mesh_terms: D030562:Databases, Protein; D045222:Electron Transport Chain Complex Proteins / Q000235:genetics / Q000378:metabolism*; D042967:Electron Transport Complex I / Q000378:metabolism; D057809:HEK293 Cells; D056945:Hep G2 Cells; D006801:Humans; D008928:Mitochondria / Q000378:metabolism*; D024101:Mitochondrial Proteins / Q000235:genetics / Q000378:metabolism*; D025941:Protein Interaction Mapping / Q000379:methods*; D060066:Protein Interaction Maps; D040901:Proteomics / Q000379:methods*; D034622:RNA Interference; D015398:Signal Transduction; D014162:Transfection; D014451:Ubiquinone / Q000378:metabolism
chemical_list: D045222:Electron Transport Chain Complex Proteins; D024101:Mitochondrial Proteins; D014451:Ubiquinone; D042967:Electron Transport Complex I
keywords: C15orf48; C2orf47; DHRS4