Role of cytoskeletal proteins in cerebral cavernous malformation signaling pathways: a proteomic analysis.
Three genetic mutations were found to cause cerebral cavernous malformation (CCM), a vascular anomaly predisposing affected individuals to hemorrhagic stroke. These CCM proteins function together as a protein complex in the cell. Loss of expression of each CCM gene results in loss of in vitro endothelial tube formation. Label-free differential protein expression analysis using multidimensional liquid chromatography/tandem mass spectrometry (2D-LC-MS/MS) was applied to explore the proteomic profile for loss of each CCM gene expression in mouse endothelial stem cells (MEES) compared to mock shRNA and no shRNA control cell-lines. Differentially expressed proteins were identified (p < 0.05). 120 proteins were differentially expressed among the cell-lines. Principal component analysis and cluster analysis show the effects of individual knockdown. In all knockdown cell-lines, altered expression of cytoskeletal proteins is the most common. While all CCM mutations result in similar pathology, different CCM mutations have their own distinct pathogenesis in cell signaling.
David H. Murdock Research Institute, North Carolina Research Campus, Kannapolis, NC 28081, USA.
Sarah Schwartz
SS
Baxter
Christopher F
CF
Dibble
Warren C
WC
Byrd
Jim
J
Carlson
Charles Russell
CR
Mack
Ivandario
I
Saldarriaga
Sompop
S
Bencharit
D000818:Animals; D002460:Cell Line; D002853:Chromatography, Liquid; D016000:Cluster Analysis; D003598:Cytoskeletal Proteins / Q000235:genetics* / Q000378:metabolism*; D042783:Endothelial Cells / Q000473:pathology / Q000502:physiology; D005786:Gene Expression Regulation; D055785:Gene Knockdown Techniques; D020786:Hemangioma, Cavernous, Central Nervous System / Q000235:genetics / Q000473:pathology*; D006801:Humans; D051379:Mice; D025341:Principal Component Analysis; D040901:Proteomics / Q000379:methods*; D015398:Signal Transduction; D053719:Tandem Mass Spectrometry
D003598:Cytoskeletal Proteins