Proteomic Identification of S-Nitrosylated Golgi Proteins: New Insights into Endothelial Cell Regulation by eNOS-Derived NO

<div><h3>Background</h3><p>Endothelial nitric oxide synthase (eNOS) is primarily localized on the Golgi apparatus and plasma membrane caveolae in endothelial cells. Previously, we demonstrated that protein S-nitrosylation occurs preferentially where eNOS is localized. Thus, in endothelial cells, Golgi proteins are likely to be targets for S-nitrosylation. The aim of this study was to identify S-nitrosylated Golgi proteins and attribute their S-nitrosylation to eNOS-derived nitric oxide in endothelial cells.</p> <h3>Methods</h3><p>Golgi membranes were isolated from rat livers. S-nitrosylated Golgi proteins were determined by a modified biotin-switch assay coupled with mass spectrometry that allows the identification of the S-nitrosylated cysteine residue. The biotin switch assay followed by Western blot or immunoprecipitation using an S-nitrosocysteine antibody was also employed to validate S-nitrosylated proteins in endothelial cell lysates.</p> <h3>Results</h3><p>Seventy-eight potential S-nitrosylated proteins and their target cysteine residues for S-nitrosylation were identified; 9 of them were Golgi-resident or Golgi/endoplasmic reticulum (ER)-associated proteins. Among these 9 proteins, S-nitrosylation of EMMPRIN and Golgi phosphoprotein 3 (GOLPH3) was verified in endothelial cells. Furthermore, S-nitrosylation of these proteins was found at the basal levels and increased in response to eNOS stimulation by the calcium ionophore A23187. Immunofluorescence microscopy and immunoprecipitation showed that EMMPRIN and GOLPH3 are co-localized with eNOS at the Golgi apparatus in endothelial cells. S-nitrosylation of EMMPRIN was notably increased in the aorta of cirrhotic rats.</p> <h3>Conclusion</h3><p>Our data suggest that the selective S-nitrosylation of EMMPRIN and GOLPH3 at the Golgi apparatus in endothelial cells results from the physical proximity to eNOS-derived nitric oxide.</p> </div

Body Design Of Tendon-Driven Jumping Robot Using Single Actuator And Wire Set

Although a mechanism in which a single actuator and a wire passing through pulleys drive the joints is a strong candidate for realizing the dynamic behavior because of its appropriate weight and simple mechanism, the problem arises that the position of the pulley influences the dynamic behavior. This paper is focused on vertical jumping. In our research, we searched an appropriate set of positions of a pulley considering the practical development of the robot and derived the relationship between the position of the pulley and the force on the tips of the robot&amp;rsquo;s foot for jumping. Simulation results suggest the possibility that some sets of positions allow an error in the attachment of the pulley, and the derived relationship indicates that the ratio of the pulling force of wire and vertical force on the ground strongly constrain the position of the pulley

Peculiar outburst of A 0535+26 observed with INTEGRAL, RXTE and Suzaku

A normal outburst of the Be/X-ray binary system A0535+26 has taken place in August 2009. It is the fourth in a series of normal outbursts that have occured around the periastron passage of the source, but is unusual by starting at an earlier orbital phase and by presenting a peculiar double-peaked light curve. A first "flare" (lasting about 9 days from MJD 55043 on) reached a flux of 440 mCrab. The flux then decreased to less than 220 mCrab, and increased again reaching 440 mCrab around the periastron at MJD 55057. Target of Opportunity observations have been performed with INTEGRAL, RXTE and Suzaku. First results of these observations are presented, with special emphasis on the cyclotron lines present in the X-ray spectrum of the source, as well as in the pulse period and energy dependent pulse profiles of the source.Comment: 6 pages, Accepted for publication on PoS, Proceedings of "The Extreme sky: Sampling the Universe above 10 keV", held in Otranto (Italy) in October 200