Identification and characterization of a novel Nogo-interacting mitochondrial protein (NIMP).
Nogo is a potent inhibitor of regeneration following spinal cord injury. To develop a better understanding of the mechanisms responsible for regenerative failure we used a yeast two-hybrid approach to try and identify proteins that interact with Nogo. We identified a novel mitochondrial protein designated Nogo-interacting mitochondrial protein (NIMP) in a screen of an adult human brain cDNA library. This interaction was confirmed by co-immunoprecipitation in both brain tissue (endogenous) and transfected HEK293T cells (overexpressed). In support of these studies we demonstrate that Nogo interacts with the UQCRC1 and UQCRC2 components of complex III, within the mitochondrial respiratory chain. The mitochondrial localization of NIMP was evidenced by confocal image analysis and western blot analysis of isolated mitochondria. NIMP is highly conserved and ubiquitously expressed in mitochondria-enriched tissues. Within the CNS, NIMP-like immunoreactivity is present in neurons and astrocytes. These data suggest that NIMP is a novel mitochondrial protein that interacts with Nogo. The interaction of Nogo with mitochondrial proteins may provide insight into the mechanisms for Nogo-induced inhibition of neurite growth.
Hu
Wen-Hui
WH
The Miami Project to Cure Paralysis, University of Miami School of Medicine, Florida 33136, USA.
Hausmann
Oliver N
ON
Yan
Ming-Shan
MS
Walters
Winston M
WM
Wong
Paul K Y
PK
Bethea
John R
JR
D000818:Animals; D019556:COS Cells; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D002417:Cattle; D002460:Cell Line; D017124:Conserved Sequence; D014450:Electron Transport Complex III / Q000378:metabolism; D006801:Humans; D046911:Macromolecular Substances; D051379:Mice; D008928:Mitochondria / Q000378:metabolism*; D024101:Mitochondrial Proteins / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D009185:Myelin Proteins / Q000378:metabolism*; D009416:Nerve Regeneration / Q000502:physiology; D000070798:Nogo Proteins; D009928:Organ Specificity; D011485:Protein Binding / Q000502:physiology; D021122:Protein Subunits; D012333:RNA, Messenger / Q000096:biosynthesis; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D020798:Two-Hybrid System Techniques
D002352:Carrier Proteins; D046911:Macromolecular Substances; D024101:Mitochondrial Proteins; D009185:Myelin Proteins; D000070798:Nogo Proteins; D021122:Protein Subunits; D012333:RNA, Messenger; C000603199:RTN4 protein, human; C459094:RTN4IP1 protein, human; D011993:Recombinant Fusion Proteins; C000603201:Rtn4 protein, mouse; D014450:Electron Transport Complex III