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Journal ArticleDOI

Proteomic characterization of the human centrosome by protein correlation profiling

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TLDR
A mass-spectrometry-based proteomic analysis of human centrosomes in the interphase of the cell cycle by quantitatively profiling hundreds of proteins across several centrifugation fractions identified and validated 23 novel components and identified 41 likely candidates as well as the vast majority of the known centrosomal proteins in a large background of nonspecific proteins.
Abstract
The centrosome is the major microtubule-organizing centre of animal cells and through its influence on the cytoskeleton is involved in cell shape, polarity and motility. It also has a crucial function in cell division because it determines the poles of the mitotic spindle that segregates duplicated chromosomes between dividing cells. Despite the importance of this organelle to cell biology and more than 100 years of study, many aspects of its function remain enigmatic and its structure and composition are still largely unknown. We performed a mass-spectrometry-based proteomic analysis of human centrosomes in the interphase of the cell cycle by quantitatively profiling hundreds of proteins across several centrifugation fractions. True centrosomal proteins were revealed by both correlation with already known centrosomal proteins and in vivo localization. We identified and validated 23 novel components and identified 41 likely candidates as well as the vast majority of the known centrosomal proteins in a large background of nonspecific proteins. Protein correlation profiling permits the analysis of any multiprotein complex that can be enriched by fractionation but not purified to homogeneity.

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Citations
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Journal ArticleDOI

Accurate Proteome-wide Label-free Quantification by Delayed Normalization and Maximal Peptide Ratio Extraction, Termed MaxLFQ

TL;DR: A new intensity determination and normalization procedure called MaxLFQ is developed that is fully compatible with any peptide or protein separation prior to LC-MS analysis, which accurately detects the mixing ratio over the entire protein expression range, with greater precision for abundant proteins.
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Locating proteins in the cell using TargetP, SignalP and related tools

TL;DR: The properties of three well-known N-terminal sequence motifs directing proteins to the secretory pathway, mitochondria and chloroplasts are described and a brief history of methods to predict subcellular localization based on these sorting signals and other sequence properties are sketched.
Journal ArticleDOI

A Model for Random Sampling and Estimation of Relative Protein Abundance in Shotgun Proteomics

TL;DR: A linear dynamic range over 2 orders of magnitude is demonstrated by using the number of spectra (spectral sampling) acquired for each protein by the data-dependent acquisition of peptides eluting into the mass spectrometer.
Journal ArticleDOI

Mass spectrometry-based proteomics turns quantitative.

TL;DR: Two recently developed methodologies offer the opportunity to obtain quantitative proteomic information by comparing the signals from the same peptide under different conditions, and stable isotope labels facilitates direct quantification from the mass spectra.
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The ABC's (and XYZ's) of peptide sequencing

TL;DR: Proteomics is an increasingly powerful and indispensable technology in molecular cell biology as discussed by the authors, which can be used to identify the components of small protein complexes and large organelles, to determine post-translational modifications and in sophisticated functional screens.
References
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Journal ArticleDOI

Mass spectrometry-based proteomics

TL;DR: The ability of mass spectrometry to identify and, increasingly, to precisely quantify thousands of proteins from complex samples can be expected to impact broadly on biology and medicine.
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Centrosome aberrations: cause or consequence of cancer progression?

TL;DR: Many human tumours show centrosome aberrations, indicating an underlying deregulation of centrosomes structure, duplication or segregation, and what impact do they have on the generation of invasive, genetically unbalanced cells during cancer progression?
Journal ArticleDOI

Centrosome composition and microtubule anchoring mechanisms.

TL;DR: The role of the centriole pair in controlling both the structural stability and the activity of theCentrosome in animal cells is now becoming clearer and the potential use of the generational asymmetry of centrosomes or spindle pole bodies for controlling cell polarity is growing theme.
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Re-evaluating centrosome function.

TL;DR: A growing body of evidence indicates that centrosomes might not be essential for spindle assembly, whereas recent data indicate that they might be important for initiating S phase and completing cytokinesis.
Journal ArticleDOI

The centrosome and cellular organization

TL;DR: The Animal Cell Centrosome and the Spindle Pole Body of Yeast Are Structurally Distinct, but Functionally Similar.
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