Protein-Glutamine Gamma-Glutamyltransferase

info:eu-repo/semantics/publishedVersio

The Redox State of Transglutaminase 2 Controls Arterial Remodeling

While inward remodeling of small arteries in response to low blood flow, hypertension, and chronic vasoconstriction depends on type 2 transglutaminase (TG2), the mechanisms of action have remained unresolved. We studied the regulation of TG2 activity, its (sub) cellular localization, substrates, and its specific mode of action during small artery inward remodeling. We found that inward remodeling of isolated mouse mesenteric arteries by exogenous TG2 required the presence of a reducing agent. The effect of TG2 depended on its cross-linking activity, as indicated by the lack of effect of mutant TG2. The cell-permeable reducing agent DTT, but not the cell-impermeable reducing agent TCEP, induced translocation of endogenous TG2 and high membrane-bound transglutaminase activity. This coincided with inward remodeling, characterized by a stiffening of the artery. The remodeling could be inhibited by a TG2 inhibitor and by the nitric oxide donor, SNAP. Using a pull-down assay and mass spectrometry, 21 proteins were identified as TG2 cross-linking substrates, including fibronectin, collagen and nidogen. Inward remodeling induced by low blood flow was associated with the upregulation of several anti-oxidant proteins, notably glutathione-S-transferase, and selenoprotein P. In conclusion, these results show that a reduced state induces smooth muscle membrane-bound TG2 activity. Inward remodeling results from the cross-linking of vicinal matrix proteins, causing a stiffening of the arterial wall

Lipid imaging of human skeletal muscle using TOF-SIMS with bismuth cluster ion as a primary ion source

Intramyocellular lipids are of importance in lipid-related diseases. The techniques in this field are limited because of a lack of adequate tools for localization of various lipids. The most usual methods for the localization of lipid distribution in the skeletal muscle are histochemistry and fluorescence probes. Different chromatography methods and mass spectrometry techniques have also been used for lipid identification. Our aim was to localize the spatial distribution of lipids in their native forms by using static time-of-flight secondary-ion mass spectrometry (TOF-SIMS). Human percutaneous skeletal muscle biopsies were obtained from the middle part of the lateral vastus muscle in the right leg of healthy adolescents with a body mass index >30. Samples were prepared by high-pressure freezing, freeze-fracturing and freeze-drying, and analysed by imaging TOF-SIMS equipped with a Bi3+ cluster ion gun. In the positive spectra, we identified phosphocholine, cholesterol, diacylglycerol, phospholipids and triacylglycerol. Phosphocholine was localized to the edge of the fibre, representing the sarcoplasma or endomysium. Weak cholesterol signals were observed in the intracellular areas. High diacylglycerol and low triacylglycerol signal intensities were seen in intracellular spaces of the transversal area of the muscle fibre. In the negative spectra, we identified fatty acids. We observed co-localization of fatty acids and diacylglycerol, which may indicate lipid-storing parts of the skeletal muscle. Thus, TOF-SIMS imaging can be used to depict the heterogeneous localization of lipids in human skeletal muscle