Plasma membrane factor XIIIA transglutaminase activity regulates osteoblast matrix secretion and deposition by affecting microtubule dynamics
| Autor: | Sarah A. Piercy-Kotb, Amina Mulani, Mari T. Kaartinen, Vamsee D. Myneni, Jeffrey W. Keillor, Hadil Al-Jallad, Nicolas Chabot |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2011 |
| Předmět: |
Macromolecular Assemblies
Anatomy and Physiology Tissue transglutaminase lcsh:Medicine Biochemistry Microtubules Cell membrane Mice 0302 clinical medicine Osteogenesis Molecular Cell Biology lcsh:Science Musculoskeletal System Cytoskeleton 0303 health sciences Multidisciplinary Factor XIII biology Chemistry Cell Differentiation Osteoblast 3T3 Cells Cellular Structures Enzymes Extracellular Matrix Cell biology medicine.anatomical_structure 030220 oncology & carcinogenesis Synthetic Chemistry Research Article 03 medical and health sciences GTP-Binding Proteins medicine Animals Protein Glutamine gamma Glutamyltransferase 2 Secretion Biology 030304 developmental biology Osteoblasts Transglutaminases Cell Membrane lcsh:R Fibronectin Tubulin Membrane protein biology.protein lcsh:Q Factor XIIIa |
| Zdroj: | PLoS ONE, Vol 6, Iss 1, p e15893 (2011) PLoS ONE |
| ISSN: | 1932-6203 |
| Popis: | Transglutaminase activity, arising potentially from transglutaminase 2 (TG2) and Factor XIIIA (FXIIIA), has been linked to osteoblast differentiation where it is required for type I collagen and fibronectin matrix deposition. In this study we have used an irreversible TG-inhibitor to 'block -and-track' enzyme(s) targeted during osteoblast differentiation. We show that the irreversible TG-inhibitor is highly potent in inhibiting osteoblast differentiation and mineralization and reduces secretion of both fibronectin and type I collagen and their release from the cell surface. Tracking of the dansyl probe by Western blotting and immunofluorescence microscopy demonstrated that the inhibitor targets plasma membrane-associated FXIIIA. TG2 appears not to contribute to crosslinking activity on the osteoblast surface. Inhibition of FXIIIA with NC9 resulted in defective secretory vesicle delivery to the plasma membrane which was attributable to a disorganized microtubule network and decreased microtubule association with the plasma membrane. NC9 inhibition of FXIIIA resulted in destabilization of microtubules as assessed by cellular Glu-tubulin levels. Furthermore, NC9 blocked modification of Glu-tubulin into 150 kDa high-molecular weight Glu-tubulin form which was specifically localized to the plasma membrane. FXIIIA enzyme and its crosslinking activity were colocalized with plasma membrane-associated tubulin, and thus, it appears that FXIIIA crosslinking activity is directed towards stabilizing the interaction of microtubules with the plasma membrane. Our work provides the first mechanistic cues as to how transglutaminase activity could affect protein secretion and matrix deposition in osteoblasts and suggests a novel function for plasma membrane FXIIIA in microtubule dynamics. |
| Databáze: | OpenAIRE |
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