Homology-based functional proteomics by mass spectrometry: application to the Xenopus microtubule-associated proteome.
The application of functional proteomics to important model organisms with unsequenced genomes is restricted because of the limited ability to identify proteins by conventional mass spectrometry (MS) methods. Here we applied MS and sequence-similarity database searching strategies to characterize the Xenopus laevis microtubule-associated proteome. We identified over 40 unique, and many novel, microtubule-bound proteins, as well as two macromolecular protein complexes involved in protein translation. This finding was corroborated by electron microscopy showing the presence of ribosomes on spindles assembled from frog egg extracts. Taken together, these results suggest that protein translation occurs on the spindle during meiosis in the Xenopus oocyte. These findings were made possible due to the application of sequence-similarity methods, which extended mass spectrometric protein identification capabilities by 2-fold compared to conventional methods.
Liska
Adam J
AJ
Max Planck Institute of Molecular Cell Biology and Genetics, Dresden, Germany.
Popov
Andrei V
AV
Sunyaev
Shamil
S
Coughlin
Peg
P
Habermann
Bianca
B
Shevchenko
Anna
A
Bork
Peer
P
Karsenti
Eric
E
Shevchenko
Andrej
A
D000818:Animals; D030562:Databases, Protein; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D008540:Meiosis / Q000502:physiology; D008869:Microtubule-Associated Proteins / Q000032:analysis* / Q000145:classification / Q000235:genetics; D008969:Molecular Sequence Data; D009865:Oocytes / Q000166:cytology / Q000502:physiology; D010802:Phylogeny; D014176:Protein Biosynthesis; D040901:Proteomics / Q000379:methods*; D008941:Spindle Apparatus / Q000737:chemistry / Q000378:metabolism / Q000648:ultrastructure; D029867:Xenopus Proteins / Q000032:analysis* / Q000145:classification / Q000235:genetics; D014982:Xenopus laevis
D008869:Microtubule-Associated Proteins; D029867:Xenopus Proteins