Volume 17, Issue 10, Pages (October 2010)

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Volume 17, Issue 10, Pages 1143-1150 (October 2010) Fluorescent Probes of Tissue Transglutaminase Reveal Its Association with Arterial Stiffening Nicolas Chabot, Simon Moreau, Amina Mulani, Pierre Moreau, Jeffrey W. Keillor Chemistry & Biology Volume 17, Issue 10, Pages 1143-1150 (October 2010) DOI: 10.1016/j.chembiol.2010.06.019 Copyright © 2010 Elsevier Ltd Terms and Conditions

Chemistry & Biology 2010 17, 1143-1150DOI: (10. 1016/j. chembiol. 2010 Copyright © 2010 Elsevier Ltd Terms and Conditions

Figure 1 TGase-Mediated Protein Crosslinking Chemistry & Biology 2010 17, 1143-1150DOI: (10.1016/j.chembiol.2010.06.019) Copyright © 2010 Elsevier Ltd Terms and Conditions

Figure 2 Rhodamine Probe 1 Used in This Work, Based on Previously Reported Irreversible Inhibitors 2 (de Macédo et al., 2002) and 3 (Pinkas et al., 2007) Chemistry & Biology 2010 17, 1143-1150DOI: (10.1016/j.chembiol.2010.06.019) Copyright © 2010 Elsevier Ltd Terms and Conditions

Figure 3 Solid-Phase Synthesis of Rhodamine Labeling Agent 1 and Control Compound 4 Chemistry & Biology 2010 17, 1143-1150DOI: (10.1016/j.chembiol.2010.06.019) Copyright © 2010 Elsevier Ltd Terms and Conditions

Figure 4 Kinetic Scheme for Irreversible Inhibition in the Presence of Substrate Chemistry & Biology 2010 17, 1143-1150DOI: (10.1016/j.chembiol.2010.06.019) Copyright © 2010 Elsevier Ltd Terms and Conditions

Figure 5 SDS-PAGE Analysis of Labeling Selectivity Incubation of activated Factor XIIIa with compound 1 (30 min, 37°C) reveals no fluorescent labeling (lane A), whereas incubation with TG2 results in strong fluorescent labeling (lane B), but not in the absence of added calcium (lane C). See Experimental Procedures for details. Chemistry & Biology 2010 17, 1143-1150DOI: (10.1016/j.chembiol.2010.06.019) Copyright © 2010 Elsevier Ltd Terms and Conditions

Figure 6 Fluorescent Staining of Aortic TG2 Staining of aortic tissues using irreversible TG2 inhibitors as fluorescent probes on control rats (A), WVK1 rats (B), WVK2 rats (C), WVK3 rats (D), and WVK4 rats (E). (F) Negative control using unreactive rhodamine derivative 7 on WVK4 rats. (G) Preincubation of compound 4 followed by incubation of compound 1 to determine the specificity of compound 1 staining on WVK4 rats. Arrows point to areas of intense TG2 labeling. A, adventitia; L, lumen; M, media of the aortic wall. Chemistry & Biology 2010 17, 1143-1150DOI: (10.1016/j.chembiol.2010.06.019) Copyright © 2010 Elsevier Ltd Terms and Conditions