Quantitative Analysis of the Human Spindle Phosphoproteome at Distinct Mitotic Stages

During mitosis, phosphorylation of spindle associated proteins is a key regulatory mechanism for spindle formation, mitotic progression, and cytokinesis. In the recent past, mass spectrometry has been applied successfully to identify spindle proteomes and phosphoproteomes, but did not address their...

Full description

Saved in:
Bibliographic Details
Published inJournal of proteome research Vol. 8; no. 10; pp. 4553 - 4563
Main Authors Malik, Rainer, Lenobel, René, Santamaria, Anna, Ries, Albert, Nigg, Erich A, Körner, Roman
Format Journal Article
LanguageEnglish
Published United States American Chemical Society 01.10.2009
Subjects
Cluster Analysis
Consensus Sequence
Culture Media
HeLa Cells
Humans
Mitosis - physiology
Phosphoproteins - analysis
Phosphorylation
Phosphoserine
Phosphothreonine
Phosphotransferases
Proteome - analysis
Reproducibility of Results
Spindle Apparatus - metabolism
Time Factors
signaling
mitosis
spindle
phosphorylation
kinase consensus
Online AccessGet full text

Cover

More Information
Summary:During mitosis, phosphorylation of spindle associated proteins is a key regulatory mechanism for spindle formation, mitotic progression, and cytokinesis. In the recent past, mass spectrometry has been applied successfully to identify spindle proteomes and phosphoproteomes, but did not address their dynamics. Here, we present a quantitative comparison of spindle phosphoproteomes prepared from different mitotic stages. In total, we report the identification and SILAC based relative quantitation of 1940 unique phosphorylation sites and find that late mitosis (anaphase, telophase) is correlated with a drastic alteration in protein phosphorylation. Further statistical cluster analyses demonstrate a strong dependency of phosphorylation dynamics on kinase consensus patterns, thus, linking subgroups of identified phosphorylation sites to known key mitotic kinases. Surprisingly, we observed that during late mitosis strong dephosphorylation occurred on a significantly larger fraction of phospho-threonine than phospho-serine residues, suggesting a substrate preference of phosphatases for phospho-threonine at this stage. Taken together, our results constitute a large quantitative data resource of phosphorylation abundances at distinct mitotic stages and they provide insight into the systems properties of phosphorylation dynamics during mitosis.
Bibliography:ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
content type line 23
ISSN:1535-3893
1535-3907
DOI:10.1021/pr9003773