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991.
S-100-Mediated Inhibition of Brain Protein Phosphorylation 总被引:7,自引:5,他引:2
Jitendra Patel Paul J. Marangos William E. Heydorn Garrick Chang Ajay Verma David Jacobowitz 《Journal of neurochemistry》1983,41(4):1040-1045
The effects of the glial-specific, calcium-binding, S-100 protein on brain membrane and supernatant protein phosphorylation were assessed. S-100 concentrations as low as 5 micrograms/ml caused a marked inhibition of the phosphorylation of a soluble brain protein having a molecular weight of 73,000 daltons (73K). This protein was designated the S-100 protein-modulated phosphoprotein (SMP). Half-maximal inhibition of the phosphorylation of SMP by S-100 was obtained at concentrations of 12 micrograms/ml (0.57 microM). The inhibition of SMP phosphorylation by S-100 was calcium-dependent, with a calculated calcium Ka of 2.0 +/- 0.3 microM. SMP phosphorylation was also inhibited by calmodulin, but only partially and with a much lower potency. The inhibition of SMP phosphorylation by S-100 was not inhibited by fluphenazine, whereas the effect of calmodulin was. SMP was found in many brain areas, with the highest levels seen in the corpus callosum. Various peripheral tissues, such as kidney; liver; and pineal, pituitary, and adrenal glands, did not contain detectable SMP levels. At higher S-100 concentrations, greater than 10 micrograms/ml, the phosphorylation of several other soluble proteins was markedly inhibited. These proteins have molecular weights of 56K, 50K, and 47K. The phosphorylation of these proteins was enhanced by calmodulin. These data suggest that the S-100 protein may function to modulate the phosphorylation of brain proteins in a manner analogous to (although in a reciprocal fashion) that of calmodulin. 相似文献
992.
Changes in the concentration of enolase isozymes and S-100 protein in degenerating and regenerating rat sciatic nerve 总被引:6,自引:4,他引:2
Levels of enolase isozymes (alpha alpha, alpha gamma, and gamma gamma forms) and S-100 protein in rat sciatic nerves were determined during their degeneration and regeneration processes. The sciatic nerves were unilaterally crushed or severed. The rats were killed 1, 2, 6, and 8-9 weeks later, and both the proximal and distal portions of the damaged nerves were dissected. Control samples were obtained from the untreated contralateral hindlimbs. Enolase isozymes and S-100 protein in the nerve segments were determined with the enzyme immunoassay method. The control nerves contained about 40, 90, and 30 pmol/mg protein of alpha alpha, alpha gamma, and gamma gamma enolases, respectively, and 0.85 microgram/mg protein of S-100 protein. These levels were not affected by repetitive electrical stimulation of the nerve fibers in vivo. The levels of the nervous system-specific forms of enolase (alpha gamma and gamma gamma) and S-100 protein decreased markedly within a week in the distal portion of the crushed nerve (alpha gamma, 27 pmol/mg; gamma gamma, 5.5 pmol/mg; S-100 protein, 0.36 microgram/mg) with apparently no change in the concentration of alpha alpha enolase. These levels in the proximal portion of the crushed nerve remained unaltered. The sensory and motor functions impaired by the sciatic nerve crush showed a recovery more or less after 4-9 weeks. This recovery was accompanied by a gradual regaining of the specific proteins in the distal portion of injured nerves (alpha gamma, 64 pmol/mg; gamma gamma, 13 pmol/mg; S-100 protein, 0.63 microgram/mg at the 8-9th week). 相似文献
993.
Bovine Brain S100 Proteins: Separation and Characterization of a New S100 Protein Species 总被引:6,自引:3,他引:3
Three S100 protein species (S100a, S100b, S100a') have been purified from bovine brain using a modification of standard preparative methods. A higher yield for each protein was obtained at the last separation step. Characterization by urea/sodium dodecyl sulfate/polyacrylamide gel electrophoresis, UV absorption spectra, and fluorescence parameters provided evidence of a new tryptophan-containing S100 protein called S100a', which exhibits, as S100a and S100b, the properties of a Ca2+ binding protein. 相似文献
994.
A rat liver cDNA library was prepared from total polyribosomal poly(A)+ RNA extracted from phenobarbital-treated animals. A cDNA clone coding for a phenobarbital-inducible cytochrome P-450 (PB P-450) was identified by differential colony hybridization to cDNAs synthesized from liver poly(A)+RNAs isolated from phenobarbital-treated rats for positive selection and cDNAs from either untreated rats or beta-naphthoflavone-treated rats as negative controls, followed by hybrid-selected translation and analysis of the translation products by immunoprecipitation. As the cloning and screening strategies involve no prior enrichment for specific mRNAs, they also permit the identification of sequences coding for phenobarbital-induced proteins other than cytochromes P-450. This relatively straightforward approach is generally applicable to the molecular cloning of sequences coding for other inducible cytochromes P-450. Nucleic acid sequencing data indicated that the cloned PB P-450 cDNA codes for a cytochrome P-450 variant [designated P-450e(U.C.)] that is very similar, but not identical, to P-450e. Sequence analysis of the section of cDNA specifying the 3'-non-coding region of the mRNA revealed that it lacked the usual poly(A) addition site signal sequence but contained three inverted repeat structures. Solution hybridization analysis demonstrated that PB P-450 mRNA is increased 20-fold by phenobarbital treatment and decreased 3-fold by beta-naphthoflavone treatment. 相似文献
995.
Blake W. Moore 《Neurochemical research》1988,13(6):539-545
The binding of Ca2+ to rat or bovine S-100 proteins, in the absence of ligands, showed a dissociation constant (in 60 mM K+) of 0.5 to 1.0 mM as measured by the effects of Ca2+ on binding of S-100 to phenyl-Sepharose, reactivity of sulfhydryl groups, and difference spectra for PHE, TYR, and TRP residues. Binding of the ligands, Stainsall and chlorpromazine lowered the dissociation constant of S-100 for Ca2+ by 2-to 10-fold as measured by the same parameters. The conformational change, in response to Ca2+ binding, probably occurs by exposure to solvent of the hydrophobic region of and subunits of S-100 at residue positions 74-93. 相似文献
996.
Summary The phenotype of Escherichia coli dnaA missense and nonsense mutations was integratively suppressed by plasmid R100-1. The suppressed strains, however, could not survive when the dnaA function was totally inactivated. This was demonstrated by the inability of replacing the dnaA allele in the suppressed strain by a dnaA::Tn10 insertion using phage P1-mediated transduction. When the intact dnaA
+ allele was additionally supplied by a specialized transducing phage, imm
21
dnaA
+, which integrated at the att
site on the E. coli chromosome, then the dnaA::Tn10 insertion, together with a oriC deletion, were able to be introduced into the suppressed strain. Thus, the mechanisms of dnaA function for oriC and for the replication origin of R100-1 may not be quite the same. 相似文献
997.
M. Mehran E. Levy M. Bendayan E. Seidman 《In vitro cellular & developmental biology. Animal》1997,33(2):118-128
Summary Although Caco-2 cells are frequently employed for the study of enterocyte lipid metabolism, variable results have been reported
regarding their ability to synthesize and secrete lipids and apolipoproteins. The major goal of this investigation is to examine
the capacity of Caco-2 cells to elaborate and secrete lipids, lipoproteins, and apolipoproteins at different degrees of morphological
and functional differentiation. Cells were cultured in medium with 5% fetal bovine serum (FBS), on permeable polycarbonate
filters from 2 to 30 d in the presence of 14C-oleate or 35S-methionine. Cellular differentiation, as assessed by morphology (light and electron microscopy), transepithelial resistance,
free fatty acid flux, and sucrase activity, progressed steadily up to 20 d of culture. Caco-2 cells esterified oleic acid
mainly into phospholipids, triglycerides (TG), and smaller amounts of cholesterol esters. Lipid synthesis began as early as
2 d, and TG secretion was enhanced with increased duration of culture. However, very low efficiency of lipid export was observed
at all levels of differentiation, reaching a maximum of only 6% of intracellular lipids. VLDL and LDL were the dominant lipoproteins
secreted, with HDL comprising <20% of the total. VLDL secretion increased, while LDL decreased, whereas the lipid composition
of lipoproteins varied little with increasing duration of culture. Apoprotein B and A-I synthesis and secretion increased
markedly from 11 to 20 d of culture. The ratio of apo B-100/B-48 decreased between 11 and 30 d, consistent with enhanced apo
B editing of more mature enterocytes. Taken together, our data suggest that from 20 d of culture, Caco-2 cells are morphologically
and functionally mature, capable of lipid esterification, and lipoprotein and apolipoprotein synthesis. However, despite their
functional and morphological similarities to mature enterocytes, Caco-2 cells have a very limited lipid export capacity. 相似文献
998.
The problem of nonspecific adsorption to the reversibly soluble-insoluble polymers is of considerable importance in the design
of an affinity precipitation protocol. It was seen that activation and coupling of the affinity ligand to the polymer changes
the nature of the polymer surface in a significant fashion. The results with pure trypsin, partially purified trypsin preparation,
and crude protein extract-containing protein inhibitors of trypsin and α-amylase and the reversibly soluble-insoluble polymer
Eudragit S-100, show that nonspecific adsorption may not be a severe limitation in such systems. 相似文献
999.
Identification and structural influence of a differentially modified N-terminal methionine in human S100b. 总被引:2,自引:2,他引:0
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S. P. Smith K. R. Barber G. S. Shaw 《Protein science : a publication of the Protein Society》1997,6(5):1110-1113
The calcium-binding protein S100b is a homodimer comprised of two identical 91-residue beta-subunits. Recombinant S100b is a heterogeneous protein, although the basis of this heterogeneity has not been established. We have used mass spectrometry and NMR spectroscopy to determine that heterogeneity in S100b arises from a mixture of formyl-S100b and desformyl-S100b when expressed in Escherichia coli. Reversed-phase HPLC purification of these two forms of S100b has allowed the differences in N-terminal composition to be used as a probe for tertiary contacts in the protein. The presence or absence of the N-terminal formyl group affected the chemical shifts of sequence neighboring residues and those in the linker of the protein (residues 40-43), indicating that these two regions are close in space. 相似文献
1000.
Identification of the binding site on S100B protein for the actin capping protein CapZ. 总被引:3,自引:1,他引:2
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P. M. Kilby L. J. Van Eldik G. C. Roberts 《Protein science : a publication of the Protein Society》1997,6(12):2494-2503
The calcium-binding protein S100B binds to several potential target proteins, but there is no detailed information showing the location of the binding site for any target protein on S100B. We have made backbone assignments of the calcium-bound form of S100B and used chemical-shift changes in spectra of 15N-labeled protein to locate the site that binds a peptide corresponding to residues 265-276 from CapZ alpha, the actin capping protein. The largest chemical-shift changes are observed for resonances arising from residues around the C terminus of the C-terminal helix of S100B and residues Val-8 to Asp-12 of the N-terminal helix. These residues are close to but not identical to residues that have been identified by mutational analysis to be important in other S100 protein-protein interactions. They make up a patch across the S100B dimer interface and include some residues that are quite buried in the structure of calcium-free S100B. We believe we may have identified a binding site that could be common to many S100 protein-protein interactions. 相似文献