caa a22 4500 477121071 CHVBK 20180405111637.0 cr unu---uuuuu 170330e19960901xx s 000 0 eng 10.1007/BF00226783 doi (NATIONALLICENCE)springer-10.1007/BF00226783 The association of phosphorylase kinase with membranes of rat liver smooth endoplasmic reticulum [Elektronische Daten] [George Maridakis, Theodore Sotiroudis] Upon fractionation of a post mitochondrial supernatant from rat liver, phosphorylase kinase activity was largely recovered in the cytosol and the smooth endoplasmic reticulum (SER) fraction. The presence of phosphorylase kinase in SER vesicles was not due to an interaction of the enzyme with glycogen particles, since previous elimination of SER glycogen either by 48 h animal starvation or by treatment of the membrane fraction with α-amylase did not significantly alter phosphorylase kinase activity content. Washing of the initial pellet of SER fraction (crude SER) by dilution and recentrifugation, released in the supernatant an amount of phosphorylase kinase activity, which is dependent on: i) the degree of dilution, ii) the number of washes, iii) the ionic strength of the washing solution and iii) the presence or absence of Ca2+. Crude SER-associated phosphorylase kinase was marginally affected by increased concentrations of antibody against rabbit skeletal muscle holoenzyme which nevertheless drastically inhibited cytosolic enzyme activity, while it showed a higher resistance to partial proteolysis and a different Western blotting profile with anti-phosphorylase kinase when compared with the soluble kinase. A small but significant fraction of SER phosphorylase kinase was strongly associated with the microsomal fraction being partly extractable only in presence of detergents. This membrane-bound enzyme form exhibited an alkaline pH optimum, in contrast to the neutral pH optima of both soluble and weakly associated phosphorylase kinase. Kluwer Academic Publishers, 1996 rat liver nationallicence phosphorylase kinase nationallicence smooth endoplasmic reticulum nationallicence SER : smooth endoplasmic reticulum nationallicence RER : rough endoplasmic reticulum nationallicence PMS : post mitochondrial supernatant nationallicence MES : 2-(N-morpholino) ethane sulfonic acid nationallicence PMSF : phenylmethylsulfonyl fluoride nationallicence SDS-PAGE : sodium dodecyl sulfate-polyacrylamide gel electrophoresis nationallicence Maridakis George Institute of Biological Research and Biotechnology, The National Hellenic Research Foundation, 48 Vassileos Constantinou Avenue, 116 35, Athens, Greece aut Sotiroudis Theodore Institute of Biological Research and Biotechnology, The National Hellenic Research Foundation, 48 Vassileos Constantinou Avenue, 116 35, Athens, Greece aut Molecular and Cellular Biochemistry Kluwer Academic Publishers; gopher//gopher.wkap.nl 154/2(1996-09-01), 153-163 0300-8177 154:2<153 1996 154 11010 https://doi.org/10.1007/BF00226783 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF00226783 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Maridakis George Institute of Biological Research and Biotechnology, The National Hellenic Research Foundation, 48 Vassileos Constantinou Avenue, 116 35, Athens, Greece aut NATIONALLICENCE 700 1- Sotiroudis Theodore Institute of Biological Research and Biotechnology, The National Hellenic Research Foundation, 48 Vassileos Constantinou Avenue, 116 35, Athens, Greece aut NATIONALLICENCE 773 0- Molecular and Cellular Biochemistry Kluwer Academic Publishers; gopher//gopher.wkap.nl/ 154/2(1996-09-01), 153-163 0300-8177 154:2<153 1996 154 11010 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer