Plasma membrane factor XIIIA transglutaminase activity regulates osteoblast matrix secretion and deposition by affecting microtubule dynamics

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 -an...

Full description

Saved in:
Bibliographic Details
Published in:PloS one 2011-01, Vol.6 (1), p.e15893-e15893
Main Authors: Al-Jallad, Hadil F, Myneni, Vamsee D, Piercy-Kotb, Sarah A, Chabot, Nicolas, Mulani, Amina, Keillor, Jeffrey W, Kaartinen, Mari T
Format: Article
Language:English
Subjects:
3T3 Cells
Animals
Biocompatibility
Biology
Cell adhesion & migration
Cell culture
Cell Differentiation
Cell Membrane - enzymology
Cell Membrane - metabolism
Cell membranes
Cell surface
Chemistry
Collagen
Collagen (type I)
Crosslinking
Cues
Dentistry
Deposition
Destabilization
Differentiation
Enzymes
Extracellular Matrix - secretion
Factor XIII - metabolism
Factor XIII - physiology
Factor XIIIa - metabolism
Fibronectin
Fibronectins
Growth factors
GTP-Binding Proteins
Immunofluorescence
Inhibition
Inhibitors
Medicine
Metabolism
Mice
Microscopy
Microtubules
Microtubules - physiology
Mineralization
Molecular weight
Osteoblastogenesis
Osteoblasts
Osteoblasts - cytology
Osteogenesis
Polymer crosslinking
Proteins
Rodents
Science
Tracking
Transglutaminase 2
Transglutaminases
Tubulin
Western blotting
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0015893