title: A protein-protein interaction map of the Trypanosoma brucei paraflagellar rod.
abstract: We have conducted a protein interaction study of components within a specific sub-compartment of a eukaryotic flagellum. The trypanosome flagellum contains a para-crystalline extra-axonemal structure termed the paraflagellar rod (PFR) with around forty identified components. We have used a Gateway cloning approach coupled with yeast two-hybrid, RNAi and 2D DiGE to define a protein-protein interaction network taking place in this structure. We define two clusters of interactions; the first being characterised by two proteins with a shared domain which is not sufficient for maintaining the interaction. The other cohort is populated by eight proteins, a number of which possess a PFR domain and sub-populations of this network exhibit dependency relationships. Finally, we provide clues as to the structural organisation of the PFR at the molecular level. This multi-strand approach shows that protein interactome data can be generated for insoluble protein complexes.
authors.affiliation: Sir William Dunn School of Pathology and Oxford Centre for Integrative Systems Biology, University of Oxford, Oxford, United Kingdom.
authors.forename: Sylvain
authors.initials: S
authors.lastname: Lacomble
authors.forename: Neil
authors.initials: N
authors.lastname: Portman
authors.forename: Keith
authors.initials: K
authors.lastname: Gull
mesh_terms: D000818:Animals; D003001:Cloning, Molecular; D000939:Epitopes; D005407:Flagella / Q000378:metabolism*; D005821:Genetic Techniques; D005822:Genetic Vectors; D023281:Genomics; D008854:Microscopy, Electron / Q000379:methods; D008954:Models, Biological; D016366:Open Reading Frames; D025941:Protein Interaction Mapping / Q000379:methods*; D040901:Proteomics / Q000379:methods; D034622:RNA Interference; D014346:Trypanosoma brucei brucei / Q000378:metabolism*; D020798:Two-Hybrid System Techniques
chemical_list: D000939:Epitopes