Has been linked to inhibition of protein translation, implicating RGS2 as a novel regulator of protein translation 103.NIH-PA Author Manuscript NIH-PA Author Manuscript NIH-PA Author Manuscript5. Functional Role of RGS Proteins in the HeartExperimental Strategies Since the discovery of RGS proteins in the heart, overexpression strategies have been used to determine the functional capacity of cardi
Kinetic formulation of the effect stands ready to be integrated into the expanding thermodynamic description of the GPCR system at large (Kenakin, 2010). From a practical point of view, the availability of naturally occurring pluridimensional effects suggests it should be possible to fine-tune the therapeutic action of a GPCR drug beyond the simple margins traditionally dictated by the target's ti
Ound in Supporting Information Fig. 6, includes eight proteins that are known to be associated with stress responses. Other proteins in this network potentially affect multiple distinct biological pathways since they are part of the translational process (Solyc01g104590.2.1, Solyc01g057830.2.1, and Solyc02g089070.2.1). Interestingly, a high proportion (64 ) of the proteins that are more abundantly
Protein characterization. iTRAQ-TAILS Validation by Glu-C Cleavage of Native Proteins--For iTRAQ-TAILS validation, we used a mixture of 13 chemokines of which one was a synthetic analog of MMPcleaved CXC motif chemokine 8 (from position 5 to 72 (5?2)) of the full-length (1?2) chemokine (supplemental Table 1). To check the accuracy of neo-N-terminal peptide identification, the chemokine mixture was
Retinopathy in animals.NIH-PA Author Manuscript NIH-PA Author Manuscript NIH-PA Author ManuscriptCytokines Busik and coworkers have presented evidence that retinal endothelial cells do not respond to hyperglycemia per se, but instead to cytokines produced by adjacent cells (Busik et al., 2008). In contrast to effects of incubating human retinal endothelial cells in high glucose, exposure of the ce
Retinopathy in animals.NIH-PA Author Manuscript NIH-PA Author Manuscript NIH-PA Author ManuscriptCytokines Busik and coworkers have presented evidence that retinal endothelial cells do not respond to hyperglycemia per se, but instead to cytokines produced by adjacent cells (Busik et al., 2008). In contrast to effects of incubating human retinal endothelial cells in high glucose, exposure of the ce
Cytoplasm in HEK293T cells (Fig. 4C). Second, we expressed NanoLuc?tagged TNIP2 (or TNIP2 Tyr230Ala as a control), together with fluorescently labeled TSG101-Halo (or unlabeled TSG101-Halo as a control), and tested for bioluminescence resonance energy transfer (NanoBRET) in HEK293T cells (42). We detected similar levels of donor luminescence for NanoLuc?TNIP2 and NanoLuc?TNIP2 Tyr230Ala samples (F
Least three independent replicates of each experiment. All of the peptide and protein information acquired in this study is provided in supplemental Table S2. Construction of -lactamase TEM-1 Fusions--N-terminal translational fusions to TEM-1 -lactamase were generated in the vector pCX341 (provided by I. Rosenshine, Hebrew University, Jerusalem, Israel) (31, 32). The coding regions, without the st