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PMID: 9585415 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Analysis of the Saccharomyces spindle pole by matrix-assisted laser desorption/ionization (MALDI) mass spectrometry.

The Journal of cell biology ·Vol. 141 ·No. 4 ·1998-05-18 ·Pages 967-77

Wigge PA, Jensen ON, Holmes S, Souès S, Mann M, Kilmartin JV

Abstract

A highly enriched spindle pole preparation was prepared from budding yeast and fractionated by SDS gel electrophoresis. Forty-five of the gel bands that appeared enriched in this fraction were analyzed by high-mass accuracy matrix-assisted laser desorption/ ionization (MALDI) peptide mass mapping combined with sequence database searching. This identified twelve of the known spindle pole components and an additional eleven gene products that had not previously been localized to the spindle pole. Immunoelectron microscopy localized eight of these components to different parts of the spindle. One of the gene products, Ndc80p, shows homology to human HEC protein (Chen, Y., D.J. Riley, P-L. Chen, and W-H. Lee. 1997. Mol. Cell Biol. 17:6049-6056) and temperature-sensitive mutants show defects in chromosome segregation. This is the first report of the identification of the components of a large cellular organelle by MALDI peptide mapping alone.

MeSH Terms
Amino Acid Sequence Chromosomes, Fungal/physiology,ultrastructure Cloning, Molecular Cytoskeletal Proteins Databases, Factual Humans Kinetochores Microscopy, Immunoelectron Models, Biological Molecular Sequence Data Mutagenesis, Site-Directed Neoplasm Proteins/chemistry Nuclear Proteins/analysis,biosynthesis,chemistry Peptide Library Peptide Mapping Saccharomyces cerevisiae/physiology,ultrastructure Saccharomyces cerevisiae Proteins Schizosaccharomyces Sequence Alignment Sequence Homology, Amino Acid Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods Spindle Apparatus/physiology,ultrastructure Temperature
Chemicals
Cytoskeletal Proteins NDC80 protein, S cerevisiae NDC80 protein, human Neoplasm Proteins Nuclear Proteins Peptide Library Saccharomyces cerevisiae Proteins
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Wigge P A
Medical Research Council (MRC) Laboratory of Molecular Biology, Cambridge CB2 2QH, United Kingdom.
Jensen O N
Holmes S
Souès S
Mann M
Kilmartin J V
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.141.4.967
Published
1998-05-18
Pages
967-77
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2132767
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.141.4.967 PMC Full Text View journal details
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