Analysis of the Saccharomyces Spindle Pole by Matrix-assisted Laser Desorption/Ionization (MALDI) Mass Spectrometry

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J. Cell Biol., Volume 141, Number 4, May 18, 1998 967-977

Analysis of the Saccharomyces Spindle Pole by Matrix-assisted Laser Desorption/Ionization (MALDI) Mass Spectrometry

Philip A. Wigge,^* Ole N. Jensen, Simon Holmes,^* Sylvie Souès,^* Matthias Mann, and John V. Kilmartin^*

^* Medical Research Council (MRC) Laboratory of Molecular Biology, Cambridge CB2 2QH, United Kingdom; and European Molecular Biology Laboratory (EMBL), D 69117 Heidelberg, Germany

A highly enriched spindle pole preparation was prepared from budding yeast and fractionated by SDS gel electrophoresis. Forty-fiveof the gel bands that appeared enriched in this fraction wereanalyzed by high-mass accuracy matrix-assisted laser desorption/ionization (MALDI) peptide mass mapping combined with sequencedatabase searching. This identified twelve of the known spindlepole components and an additional eleven gene products that hadnot previously been localized to the spindle pole. Immunoelectronmicroscopy localized eight of these components to different partsof the spindle. One of the gene products, Ndc80p, shows homologyto human HEC protein (Chen, Y., D.J. Riley, P-L. Chen, and W-H.Lee. 1997. Mol. Cell Biol. 17:6049-6056) and temperature-sensitivemutants show defects in chromosome segregation. This is the firstreport of the identification of the components of a large cellularorganelle by MALDI peptide mapping alone.

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