Proteomic analysis of mammalian primary cilia.

Abstract

The primary cilium is a microtubule-based organelle that senses extracellular signals as a cellular antenna. Primary cilia are found on many types of cells in our body and play important roles in development and physiology. Defects of primary cilia cause a broad class of human genetic diseases called ciliopathies. To gain new insights into ciliary functions and better understand the molecular mechanisms underlying ciliopathies, it is of high importance to generate a catalog of primary cilia proteins. In this study, we isolated primary cilia from mouse kidney cells by using a calcium-shock method and identified 195 candidate primary cilia proteins by MudPIT (multidimensional protein identification technology), protein correlation profiling, and subtractive proteomic analysis. Based on comparisons with other proteomic studies of cilia, around 75% of our candidate primary cilia proteins are shared components with motile or specialized sensory cilia. The remaining 25% of the candidate proteins are possible primary cilia-specific proteins. These possible primary cilia-specific proteins include EVC2, INPP5E, and inversin, several of which have been linked to known ciliopathies. We have performed the first reported proteomic analysis of primary cilia from mammalian cells. These results provide new insights into primary cilia structure and function.

• Authors: Hiroaki Ishikawa, James Thompson, John R Yates 3rd, Wallace F Marshall
• Journal: Current biology : CB (Curr Biol) Vol. 22 Issue 5 Pg. 414-9 (Mar 06 2012) ISSN: 1879-0445 [Electronic] England
• PMID: 22326026 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2012 Elsevier Ltd. All rights reserved.

Chemical References

• EVC2 protein, mouse
• Intercellular Signaling Peptides and Proteins
• Invs protein, mouse
• Membrane Proteins
• Proteome
• Transcription Factors
• Phosphoric Monoester Hydrolases
• phosphoinositide 5-phosphatase

Topics

• Animals
• Cilia (chemistry, physiology, ultrastructure)
• Intercellular Signaling Peptides and Proteins
• Kidney (cytology, physiology)
• Membrane Proteins (isolation & purification)
• Mice
• Phosphoric Monoester Hydrolases (isolation & purification)
• Protein Array Analysis (methods)
• Proteome (chemistry, physiology)
• Proteomics (methods)
• Transcription Factors (isolation & purification)