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181.
A Akatsuka T J Singh H Nakabayashi M C Lin K P Huang 《The Journal of biological chemistry》1985,260(6):3239-3242
Addition of glucagon (20 nM) to the isolated hepatocytes from 24-h starved male rats results in an inactivation of glycogen synthase. The A0.5 for glucose-6-P is increased 2-fold over the control but the S0.5 for UDP-glucose is not significantly affected. The glucagon-stimulated inactivation of glycogen synthase is also accompanied by a 60-120% increase in the phosphorylation of the synthase. Glycogen synthase labeled with 32P by incubation of the hepatocytes with [32P] PO4(3-) was recovered by immunoprecipitation and the resulting immunoprecipitate was subjected to tryptic digestion. Analysis of the 32P-labeled peptides reveals that the sites corresponding to those phosphorylated by cAMP-dependent protein kinase and glycogen synthase (casein) kinase-1 (Itarte, E., and Huang, K.-P. (1979) J. Biol. Chem. 254, 4052-4057) are rapidly phosphorylated in response to glucagon. These results demonstrate that glucagon not only triggers the activation of cAMP-dependent protein kinase through an increase in the intracellular level of cAMP but also, by an unknown mechanism, activates a Ca2+- and cAMP-independent protein kinase. 相似文献
182.
Paul P. C. Lin 《Plant Growth Regulation》1985,3(3-4):257-268
Incorporation of L-[U-14C] arginine or L-[U-14C] ornithine into putrescine (Put), spermidine (Spd) and spermine (Spm) in embryonectomized barley seeds (Hordeum vulgare L. cv. Himalaya) was studied following imbition with methylglyoxal-bis (guanylhydrazone) (MGBG) and abscisic acid (ABA). Both radiolabeled amino acids were incorporated into the amines as a result of active polyamine biosynthesis in the seed during imbibition. In the aleurone layer, the Spd and Spn existed mainly in the free form (acid soluble). However about 50% of Put was recovered in conjugated form(s) (acid insoluble). Imbibition with 5 and 10M ABA for 3 days increased the accumulation of the free form of 14C-Put, probably as a result of inhibition (70%) of 14C-Spd accumulation. The ABA treatment showed no significant effect on levels of the conjugated form of Put and Spd. Imbibition with millimolar concentrations of MGBG resulted in (i) abnormal accumulation of the free form of Put and incorporation of 14C-amino acids into the diamine, (ii) progressive inhibition of the accumulation of the free forms of 14C-Spd and Spm, and (iii) reduction of the 14C incorporation into the conjugated forms of Put and Spd. Uptake of 14C-amino acids was not affected by MGBG treatment. The results indicate that MGBG may inhibit not only the synthesis of Spd and Spm, but the catabolism (e.g. oxidation) of Put in the aleurone layer.This paper is published with the approval of the director of the Kentucky Agricultural Experiment Station. 相似文献
183.
C W Chi D X Zhu N Q Lin L X Xu F L Tan L X Wang 《Biological chemistry Hoppe-Seyler》1985,366(9):879-885
After reduction and alkylation of the disulfide bonds of the proteinase inhibitor B from the root of the arrowhead (Sagittaria sagittifolia L.) followed by CNBr cleavage three peptide fragments with 68, 62 and 11 amino-acid residues could be separated on DEAE-Sepharose CL-6B. The peptides or the inhibitor itself were further specifically cleaved either by trypsin or by the mixture of (CH3)2SO/HCl/HBr at the arginyl- and the tryptophyl-peptide bond, respectively. The complete amino-acid sequences of the peptides were determined by manual solid phase DABITC/PITC double coupling micro-method and the primary structure of the arrowhead inhibitor B consisting of 141 amino-acid residues was then elucidated. Twenty pairs of amino-acid residues are repeated in the molecule of this inhibitor, three of these pairs even occur three times. The possible locations of the reactive sites are discussed. On the basis of sequence comparisons between this inhibitor and all other serine proteinase inhibitors the arrowhead inhibitor may belong to a new family. 相似文献
184.
Summary Microwave irradiation has been shown to decrease the input resistance of snail neurons. In this study, we examined the role of extracellular calcium in triggering the microwave-induced enhancement of membrane conductance. Two sets of experiments were conducted. In the first set, nerve cells were superfused using Ringer solution with added Cd2+ (0.9 mM) which is a known blocker of calcium channels. In the second set, cells were superfused with low Ca2+ (0.7 mM) Ringer solution. Microwave irradiation was conducted at 2,450 MHz for 30 min with a specific absorption rate of 13 mW/g. It was found that 7 mM to 0.7 mM lowering of Ca2+ in bathing solution as well as blocking of calcium channels in neuronal membrane by means of Cd2+ did not influence the fall in membrane resistance induced by microwave radiation. In fact, the observed changed in membrane resistance in these experiments were nearly equal to those observed for neurons superfused by normal Ringer's. Thus, these results rule out the possible contribution of external Ca2+ in the observed microwave effect. Experiments with high Ca2+ solution also support this conclusion. 相似文献
185.
Synthesis and phosphorylation of cytoskeleton components in foetal,regenerating and adult normal rat hepatocytes during culture 总被引:1,自引:0,他引:1
Georges Baffet Annie Ruelland Bruno Clement Elisabeth Le Rumeur Siegmund Fischer 《Molecular and cellular biochemistry》1985,68(2):97-105
Summary Detergent insoluble material (DIM) was prepared by gentle treatment with detergent from foetal, regenerating and adult normal rat hepatocytes cultured for various times. It retained to some degree the morphology of the cells. After incubation of intact cells with 35S-methionine, most of the labelled DIM proteins were found to be components of the cytoskeleton. They included several cytokeratins, vimentin and actin. The synthesis rate varied with the age of animals and culture conditions. The high synthetic rate of vimentin in foetal and regenerating hepatocytes could be associated with cell proliferation. No correlation was found between cytokeratin synthesis and hepatocyte growth. Most of the cytoskeleton proteins could be phosphorylated in intact cells and in DIM from cultured hepatocytes. However the degree of phosphorylation of these proteins was not related to their synthetic rate. The decreased phosphorylation level in cultured adult rat hepatocytes could be related to the rapid loss of specific functions. 相似文献
186.
Cloning of the pectate lyase genes from Erwinia carotovora and their expression in Escherichia coli 总被引:11,自引:0,他引:11
A hybrid cosmid coding for pectate lyase (PL) activity was identified from an Erwinia carotovora genomic library by an immunological screening method. A 7-kb DNA fragment was identified which codes for three proteins identical in size to proteins with PL activity purified from E. carotovora culture supernatants. The three proteins had apparent Mrs of 41, 44 and 44 X 10(3) as estimated by SDS-PAGE. None of the PLs were exported from Escherichia coli strain HB101 but all were found in the periplasmic space. Plant tissue was macerated by the PLs made in E. coli. 相似文献
187.
The araBAD operon of Salmonella typhimurium LT2. I. Nucleotide sequence of araB and primary structure of its product, ribulokinase 总被引:6,自引:0,他引:6
Hybrid plasmids containing the araBAD operon of Salmonella typhimurium LT2 were characterized by Southern blot and genetic analyses. The nucleotide sequence of araB was determined. The araB gene product, ribulokinase (EC 2.7.1.16), was purified and the results of amino acid composition analysis and partial amino acid sequence are in agreement with predictions from the DNA sequence. Ribulokinase is 569 amino acid residues long and has a calculated Mr of 61 793. Ribulokinase shares significant homology with xylulose kinase from Escherichia coli. Codon usage in the araB gene does not favor those codons which have intermediate codon-anticodon binding energy. 相似文献
188.
189.
190.
武汉东湖浮游植物各种成份分析与沉淀物中浮游植物活体碳、氮、磷的测定 总被引:4,自引:2,他引:2
本实验调查了武汉东湖浮游植物水华的各种成份以及水柱沉淀物内叶绿素α含量,根据碳与叶绿素α关系,给定了一回归方程,并计算出沉淀物中藻类活体相应的碳、氮、磷含量。从平均数值看,武汉东湖浮游植物水华的C,N,P含量分别为39.30,7.98及0.94(%);其干湿比为0.20,碳、氮比为5.10,碳、磷比为46.54,碳与叶绿素α之比则为133.32。 1983年东湖水柱沉淀物中浮游植物活体叶绿素α下沉量平均每天每平方米为43.8675微克(Ⅰ站)及35.5881微克(Ⅱ站)。利用东湖浮游植物水华各种成份含量及其各种比率计算出的东湖水柱沉淀物中浮游植物活体碳、氮、磷量,按顺序每天每平方米分别为2.53,0.50,0.05毫克(Ⅰ站)及2.09,0.41,0.04毫克(Ⅱ站)。 相似文献