Proteome analysis of microtubule-associated proteins and their interacting partners from mammalian brain.
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.
Kozielski
Frank
F
The Beatson Institute for Cancer Research, Switchback Road, Bearsden, Glasgow, G61 1BD, Scotland, UK.
Riaz
Tahira
T
DeBonis
Salvatore
S
Koehler
Christian J
CJ
Kroening
Mario
M
Panse
Isabel
I
Strozynski
Margarita
M
Donaldson
Ian M
IM
Thiede
Bernd
B
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*
D010750:Phosphoproteins; D020033:Protein Isoforms; D020543:Proteome; D014404:Tubulin