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41.
Robert K. MacNeal Bradford C. Webb Daniel L. Purich 《Biochemical and biophysical research communications》1977,74(2):440-447
Tubulin derived from cold depolymerized bovine microtubules has been gel filtered to obtain a tubulin preparation with only 3% of the tubulin dimers containing exchangeable [3H]-guanine nucleotide. In the presence of acetyl-P and bacterial acetate kinase, this preparation polymerizes to form microtubules which are morphologically indistinguishable from microtubules formed in the presence of excess GTP. The extent of microtubule formation at substoichiometric nucleotide levels using the GTP regenerating system exceeds the extent of assembly obtained with excess GTP. It is concluded that the exchangeable guanine nucleotide site can be virtually unoccupied in intact neurotubules and this finding indicates that GDP can “catalyze” tubule assembly in the presence of a GTP regenerating system. 相似文献
42.
Seventeen male crab-eating macaques, drawn from two captive troops, were tested on a series of complex problem solving tasks
in a Wisconsin General Test Apparatus (wgta). The animals were trained on a series of 6-trial object quality learning set problems followed by a series of 10-trial object
quality learning set problems. They were then given problems in which the correct stimulus object was reversed part way through
the problem. After the animals reached criterion on this task, the reversal learning set was then extinguished. High ranking
animals made more intraproblem errors than low ranking animals on the 6-trial problems, but there was no relationship between
social status and the rapidity with which the object quality learning set was established. Animals that received overtraining
on the 6-trial problems transferred their learning virtually intact to the 10-trial problems; however, high ranking animals
without overtraining made more errors than low ranking animals. On reversal learning and reversal extinction, high ranking
animals made more errors on critical trials, indicating that they formed and extinguished the reversal set more slowly than
low ranking animals. Object quality sets, as measured by trial-2 performance, were not affected by the reversal conditions.
Supported by USAMRDC Contract No. DADA 17-73-C-3007. 相似文献
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An insulin-stimulated kemptide kinase purified from rat liver is deactivated by phosphatase 2A. 总被引:1,自引:0,他引:1
J K Klarlund S R Jaspers N Khalaf A P Bradford T B Miller M P Czech 《The Journal of biological chemistry》1991,266(7):4052-4055
Insulin action leads to the rapid stimulation of a cytosolic Kemptide (Leu-Arg-Arg-Ala-Ser-Leu-Gly) kinase (KIK) that has been recently purified to near homogeneity (Klarlund, J. K., Bradford, A. P., Milla, M. G., and Czech, M. P. (1990) J. Biol. Chem. 265, 227-234). To examine its activation mechanism, purified KIK was treated with purified protein phosphatases. The catalytic subunit of phosphatase 2A inhibited the activity of control KIK by about 50% and abolished the 5-fold elevation in KIK activity due to insulin action. The catalytic subunit of phosphatase 1 with equivalent activity based on dephosphorylation of 32P-labeled phosphorylase alpha had no effect on either control or insulin-stimulated KIK activity. The deactivation of insulin-stimulated KIK by phosphatase 2A was time- and concentration-dependent and was blocked by phosphatase inhibitors. The purified native complexes of phosphatase 2A, phosphatase 2A1, and phosphatase 2A2 similarly deactivated KIK. Analyis of control or insulin-stimulated KIK with two antiphosphotyrosine antibodies by immunoblotting and immunoprecipitation failed to detect the presence of phosphotyrosine in the kinase. These results indicate that KIK is activated by phosphorylation as part of a kinase cascade emanating from insulin receptor stimulation. 相似文献
46.
A M Irani S M Goldstein B U Wintroub T Bradford L B Schwartz 《Journal of immunology (Baltimore, Md. : 1950)》1991,147(1):247-253
Two murine mAb were prepared against human mast cell carboxypeptidase (HMC-CP) purified from human skin, and were termed CP1 and CP2, respectively. Double immunohistochemical labeling of Carnoy's-fixed sections of human skin, lung, and gastrointestinal tissue with CP1 and CP2, respectively, and with a murine monoclonal antitryptase antibody demonstrated that HMC-CP was selectively present in a subset of human mast cells. Double labeling experiments with CP1 and CP2, respectively, and a murine anti-chymase mAb demonstrated the presence of HMC-CP in the tryptase-positive, chymase-positive mast cell type (MCTC) only. Immunohistochemical labeling of peripheral blood leukocytes resulted in staining of monocytes with CP2 but not with CP1. In addition to chymase and a cathepsin-G like proteinase, HMC-CP is another neutral protease that is selectively present in the MCTC tryptase-positive, chymase-positive mast cells type of mast cell, thus extending the biochemical definition of human mast cell heterogeneity. 相似文献
47.
L Wolfe A P Bradford J K Klarlund M P Czech 《The Journal of biological chemistry》1992,267(14):9749-9756
A cytosolic insulin-sensitive serine kinase has been purified to apparent homogeneity in parallel from livers of control or acutely insulin-treated rats. The kinase is labile and requires rapid purification for stability. The kinase migrates as a band of apparent Mr = 90,000 on denaturing gels and elutes as a monomer on Superose 12 gel filtration. After sodium dodecyl sulfate-polyacrylamide gel electrophoresis and renaturation, the 90-kDa band presumed to be the kinase shows kinase activity toward myelin basic protein in situ. Substrates of the kinase include Leu-Arg-Arg-Ala-Ser-Leu-Gly (Kemptide), ribosomal protein S6, S6 peptide, a proline-rich peptide substrate, microtubule-associated protein 2, and myelin basic protein. The kinase also phosphorylates histones H1 and H2B, but does not autophosphorylate to a significant stoichiometry. The activity of the kinase is inhibited by fluoride, glycerophosphate, p-nitrophenyl phosphate, p-nitrophenol, heparin, quercetin, poly-L-lysine, and potassium phosphate, but is unaffected by calcium, cAMP, spermine, protein kinase inhibitor peptide, phorbol myristate acetate, calcium plus phosphatidylserine, or vanadate. The kinase will utilize magnesium (10 mM) as well as manganese (1 mM) as a cofactor for maximal phosphotransferase activity. The kinase is not detected by immunoblotting with antibodies directed against protein kinase C or type II S6 kinase. Taken together, these properties distinguish this kinase from other insulin-sensitive kinases that have been described previously. The purified kinase from livers of insulin-treated rats shows a 5-20-fold higher specific activity compared to enzyme prepared from control rats, suggesting a covalent modification as the mechanism of activation. Incubation of purified, insulin-stimulated kinase with purified phosphatase 2A leads to deactivation of the kinase activity, and the phosphatase inhibitor nitrophenyl phosphate blocks this deactivation. The insulin-activated kinase fails to immunoblot with anti-tyrosine phosphate antibodies. Taken together, these results indicate that insulin activates this novel cytosolic protein kinase by a mechanism that causes its phosphorylation on serine or threonine residues. 相似文献
48.
49.
Occurrence of Vibrio cholerae serotype O1 in Maryland and Louisiana estuaries. 总被引:36,自引:27,他引:9 下载免费PDF全文
R R Colwell R J Seidler J Kaper S W Joseph S Garges H Lockman D Maneval H Bradford N Roberts E Remmers I Huq A Huq 《Applied microbiology》1981,41(2):555-558
Vibrio cholerae serotype O1 has been isolated from Chesapeake Bay in Maryland and estuaries and sewers in Louisiana. The occurrence of V. cholerae O1 in the aquatic environment in the absence of human disease suggests that this organism survives and multiples in the natural environment. 相似文献
50.
Influence of Protein Synthesis on NO(3) Reduction, NH(4) Accumulation, and Amide Synthesis in Suspension Cultures of Paul's Scarlet Rose 下载免费PDF全文
Changes in the concentrations of NH4+ and amides during the growth of suspension cultures of rose (Rosa cv. Paul's Scarlet) cells were examined. When cells were grown in medium possessing only NO3− as a nitrogen source, the concentrations of NH4+ and amides increased to 4.0 × 10−1 and 5.9 micromoles per gram fresh weight, respectively. The amounts of both constituents declined during the later stages of growth. When a trace amount of NH4+ was added to the NO3− base starting medium, the concentration of NH4+ in the cells was increased to 7.0 × 10−1 micromoles per gram fresh weight. 相似文献