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41.
Hirofumi Ogawa Yoshihisa Ishiguro Motoji Fujioka 《Archives of biochemistry and biophysics》1983,226(1):265-275
Guanidoacetate methyltransferase (EC 2.1.1.2) has been purified about 800-fold from rat liver. The purified preparation shows a single protein band on polyacrylamide gel electrophoresis in the presence and absence of sodium dodecyl sulfate. The molecular weight of the enzyme is estimated to be 25,000 and 26,000 by Sephadex gel molecular-exclusion chromatography and by electrophoresis in polyacrylamide gradient gel, respectively. The sodium dodecyl sulfate-denatured enzyme also has a molecular weight of 26,000; thus, the enzyme is a monomeric protein. Guanidoacetate methyltransferase as isolated is catalytically inactive, but is readily reactivated by incubation with a thiol. The reactivated enzyme, which contains 3 mol of sulfhydryl groups/mol of enzyme, is again inactivated by oxidized glutathione. This inactivation is accompanied by the disappearance of two sulfhydryl residues. The relationship between the loss of enzyme activity and the number of residues disappeared indicates that the integrity of these sulfhydryl residues is critical for activity. The oxidized enzyme fails to bind the substrate S-adenosylmethionine as evidenced by the equilibrium dialysis study. Alkylation of the nonoxidizable sulfhydryl by N-ethylmaleimide shows that this residue is also essential for activity. UV absorption, fluorescence, and CD spectra show no difference between the reduced and oxidized enzymes, but the former is more susceptible to proteolytic attack by trypsin. The enzyme has an isoelectric pH of 5.3, and is most active at pH 9.0. From the CD spectrum, an α helix content of 15% is calculated. The Km values for guanidoacetate and S-adenosylmethionine are 97.5 and 6.73 μm, respectively, at pH 8.0 and 37 °C. 相似文献
42.
Isolation of virus causing hemorrhagic fever with renal syndrome (HFRS) through a cell culture system 总被引:4,自引:0,他引:4
T Kitamura C Morita T Komatsu K Sugiyama J Arikawa S Shiga H Takeda Y Akao K Imaizumi A Oya N Hashimoto S Urasawa 《Japanese journal of medical science & biology》1983,36(1):17-25
Twenty-three rat lung specimens collected in outbreaks of hemorrhagic fever with renal syndrome (HFRS) in three medical institutions were inoculated onto the VERO-E6 cell monolayers. After several blind passages, an agent growing serially in the cell cultures and reacting specifically with known HFRS-positive sera was isolated from two of these specimens. The two isolates were antigenically identical each other. The agent, named strain SR-11, was identified as the causative virus of HFRS by its antigenic identity with E6 cell-adapted HFRS virus, Hantaan 76-118 strain, and the specific reactions with sera from various HFRS cases. 相似文献
43.
Takeda Junko; Abe Shunnosuke; Morikawa Hiromichi; Senda Mitsugi 《Plant & cell physiology》1983,24(4):667-676
The electrophysiological properties of the membrane of Nicotianatabacum var. Sarnsun cultured cells were determined using amicroelectrode technique in standard medium containing 1 mMKC1, 1 mM NaCl and 1 mu CaCl2 at pH 7. Tobacco callus was derivedfrom the pith (Em=104.4%16.2 mV). The membrane potentialsof the callus cells did not show a symmetrical Gaussian distributionbut were scattered over a wide range. The percentage of highmembrane potential cells increased as the subculture was continueduntil about 11 months and then decreased. The response of themembrane potential to electric stimulus, ionic composition,metabolic inhibitors, sugars and amino acids was characteristicof high (Em={small tilde}160 220 mV; H-cells)and low (Em=80{small tilde}90 mV; L-cells) membranepotential cells. The membrane potential of H-cells was largelydepolarized by addition of CN, carbonium cyanide m-chlorophenylhydrazone,decyclohexylcarbodiimide, and triphenyltin chloride and transientlydepolarized by addition of glucose, galactose, mannose or sucrose,and D-alanine, L-alanine or Llysine, but the membrane potentialof L-cells was not. (Received December 3, 1982; Accepted March 16, 1983) 相似文献
44.
Nuclear phosphoprotein kinases from rat liver 总被引:3,自引:0,他引:3
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46.
Studies on ATP citrate lyase of rat liver. IV. The role of CoA 总被引:1,自引:0,他引:1
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48.
A thermolabile direct hemolysin of Vibrio parahaemolyticus 总被引:6,自引:0,他引:6
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