Quantitative proteomics reveals the protective effects of ESD against osteoarthritis via attenuating inflammation and modulating immune response

Epimedium, Salvia miltiorrhiza, and Dioscorea nipponica Makino (ESD) have been combined to treat osteoarthritis (OA) for a long time. In this study we used quantitative proteomics to find the protective effects of ESD against OA and possible mechanism. After papain-induced rats’ OA model established ESD was intragastrically administrated to rats for four weeks. Label-free quantitative proteomics was used to screen the comprehensive protein profiling changes in both OA and ESD groups. After stringent filtering, 62 proteins were found to be significantly up-regulated and 208 proteins were down-regulated in OA model compared with sham-operated control. Functional analysis revealed that these OA up-regulated proteins were enriched in the activation of humoral immunity response, complement activation, leukocyte mediated immunity, acute inflammatory, endocytosis regulation, and proteolysis regulation. ESD partially recovered the protein profiling changes in OA model. The effects of ESD were also assessed by measurement of behavioral activity and pathologic changes in the joints. ESD showed protective effects in suppressing inflammation, releasing joint pain, and attenuating cartilage degradation. Our study presented that ESD as a potential candidate to alleviate OA damage by reducing inflammation and modulating of immune system.

Involvement of Regucalcin in Protein Synthesis and Proteolysis: Regulation of the Enzyme Activity in the Cytoplasm and Nucleus

<p>Regucalcin was discovered in 1978 as a novel calcium-binding protein. The name, regucalcin, was proposed for this calcium-binding protein, which regulates various Ca^2+- or Ca²⁺/calmodulin-dependent enzyme activations. The regucalcin gene (gene symbol;rgn) is localized on the X chromosome. Regucalcin has been demonstrated to play a multifunctional role in the regulation of intracellular calcium homeostasis, signal transduction, gene expression, cell proliferation and apoptosis in various types of cells and tissues. The cytoplasmic regucalcin translocases to the nucleus and suppresses nuclear deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) synthesis. Moreover, regucalcin has been shown to reveal a suppressive effect on protein synthesis and a stimulatory effect on protein degradation. Regucalcin has been found to inhibit aminoacyl-tRNA synthetase and activate thiol protease. Â Regucalcin may play a suppressive role in the regulation of protein turnover in cells.</p>

The two PAN ATPases from Halobacterium display N-terminal heterogeneity and form labile complexes with the 20S proteasome

The PAN (proteasome-activating nucleotidase) proteins from archaea represent homologues of the eukaryotic 26S proteasome regulatory ATPases. In vitro the PAN complex has been previously shown to have a stimulatory effect on the peptidase activities of the 20S core. By using gradient ultracentrifugation we found that, in cellular extracts, the two PAN proteins from Halobacterium do not form stable high-molecular-mass complexes. Only PAN B was found to associate transiently with the 20S proteasome, thus suggesting that the two PAN proteins are not functionally redundant. The PAN B–20S proteasome complexes associate in an ATP-dependent manner and are stabilized upon nucleotide binding. The two PAN proteins were immunodetected in cellular extracts as N-terminal-truncated polypeptides. RNA-mapping experiments and sequence analysis indicated that this process involved transcript heterogeneities and dual translational initiation mechanisms. Taken together, our results suggest that PAN N-terminal modifications and their intracellular dynamics of assembly/association may constitute important determinants of proteolysis regulation.

Cell hydration controls autophagosome formation in rat liver in a microtubule-dependent way downstream from p38MAPK activation

Autophagic proteolysis in rat liver is under the control of the cellular hydration state. Because the morphological site of swelling-dependent proteolysis regulation has not yet been identified, the formation of autophagosomes was investigated with transmission electron microscopy in slices from perfused livers. In livers from fed rats, hypo-osmotic exposure (185mosmol/l) led within 30min to a decrease in fractional cytoplasmic autophagosome volume that was sensitive to colchicine and p38MAPK inhibition. Similarly, the decrease in autophagosome volume, but not the increase in cell volume caused by insulin or glutamine/glycine, was strongly inhibited by colchicine and SB 203580, an inhibition of p38MAPK activation. Immune complex assays from perfused liver showed that hypo-osmotic activation of p38MAPK was not inhibited by colchicine. Further, experiments using confocal laser microscopy in cultivated hepatocytes incubated with mouse-derived anti-(α-tubulin) showed that microtubular structures were not influenced by the inhibition of p38MAPK by SB 203580. It is concluded that the sequestration of autophagic vacuoles is a major site of proteolysis regulation by cell hydration. Swelling-induced activation of p38MAPK is required for this process and occurs upstream of the putative microtubule regulation site.