title: Proteome analysis of microtubule-associated proteins and their interacting partners from mammalian brain.
abstract: The microtubule (MT) cytoskeleton is essential for a variety of cellular processes. MTs are finely regulated by distinct classes of MT-associated proteins (MAPs), which themselves bind to and are regulated by a large number of additional proteins. We have carried out proteome analyses of tubulin-rich and tubulin-depleted MAPs and their interacting partners isolated from bovine brain. In total, 573 proteins were identified giving us unprecedented access to brain-specific MT-associated proteins from mammalian brain. Most of the standard MAPs were identified and at least 500 proteins have been reported as being associated with MTs. We identified protein complexes with a large number of subunits such as brain-specific motor/adaptor/cargo complexes for kinesins, dynein, and dynactin, and proteins of an RNA-transporting granule. About 25% of the identified proteins were also found in the synaptic vesicle proteome. Analysis of the MS/MS data revealed many posttranslational modifications, amino acid changes, and alternative splice variants, particularly in tau, a key protein implicated in Alzheimer's disease. Bioinformatic analysis of known protein-protein interactions of the identified proteins indicated that the number of MAPs and their associated proteins is larger than previously anticipated and that our database will be a useful resource to identify novel binding partners.
authors.affiliation: The Beatson Institute for Cancer Research, Switchback Road, Bearsden, Glasgow, G61 1BD, Scotland, UK.
authors.forename: Frank
authors.initials: F
authors.lastname: Kozielski
authors.forename: Tahira
authors.initials: T
authors.lastname: Riaz
authors.forename: Salvatore
authors.initials: S
authors.lastname: DeBonis
authors.forename: Christian J
authors.initials: CJ
authors.lastname: Koehler
authors.forename: Mario
authors.initials: M
authors.lastname: Kroening
authors.forename: Isabel
authors.initials: I
authors.lastname: Panse
authors.forename: Margarita
authors.initials: M
authors.lastname: Strozynski
authors.forename: Ian M
authors.initials: IM
authors.lastname: Donaldson
authors.forename: Bernd
authors.initials: B
authors.lastname: Thiede
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001921:Brain / Q000378:metabolism*; D002417:Cattle; D006801:Humans; D008870:Microtubules / Q000378:metabolism*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010750:Phosphoproteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D025941:Protein Interaction Mapping; D020033:Protein Isoforms / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D020543:Proteome / Q000378:metabolism*; D053719:Tandem Mass Spectrometry; D014404:Tubulin / Q000378:metabolism*
chemical_list: D010750:Phosphoproteins; D020033:Protein Isoforms; D020543:Proteome; D014404:Tubulin