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61.
Phosphorylation and dephosphorylation of ribosomal proteins have been suggested to participate in the regulation of protein synthesis in eukaryotic organisms. The present research focuses on the purification and partial characterization of a protein kinase from maize ribosomes that specifically phosphorylates acidic ribosomal proteins. Ribosomes purified from maize axes were used as the enzyme source. Purification of ribosomes was performed by centrifugation through a 0.5 M sucrose, 0.8 M KCl cushion. A protein kinase activity present in this fraction was released by extraction with 1.5 M KCl and further purified by diethylaminoethyl cellulose column chromatography. A peak containing protein kinase activity was eluted around 400 m M KCl. Analysis of this fraction by sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed one band of 38 kDa molecular mass, which cross-reacted in a western blot with antibodies raised against proteins from the large ribosomal subunit. This enzyme specifically phosphorylates one of the acidic ribosomal proteins (P2 ). Its activity is inhibited by Ca2+ and Zn2+ and is activated by Mg2+ , polylysine and spermine. The relevance of this protein kinase in reinitiating the protein synthesis process during germination is discussed. 相似文献
62.
Manuel B. Aguilar Daniel Soyez Rocco Falchetto David Arnott Jeffrey Shabanowitz Donald F. Hunt Alberto Huberman 《Peptides》1995,16(8):1375-1383
The primary structure of the neurohormone crustacean hyperglycemic hormone (CHH-II) was determined by means of enzymatic digestions, manual Edman degradation, and mass spectrometry. CHH-II is a 72 residue peptide (molecular mass 8388 Da), with six cysteines forming three disulfide bridges that connect residues 7–43, 23–39, and 26–52. The peptide has blocked N- and C-termini, and lacks tryptophan, histidine, and methionine. The CHH-I and CHH-II of Procambarus bouvieri have identical sequences and elicit levels of hyperglycemia that are not distinguishable. The difference between the two isomorphs consists in a posttranslational modification of a l-Phe in CHH-I to a d-Phe in CHH-II at the third position from the N-terminus. 相似文献
63.
A simple linear relationship between the J
coupling constant and the linewidth (1/2) of in-phase NMR peaks has been identified. This relationship permits the rapid and accurate determination of polypeptide J
coupling constants from a simple inspection of amide cross peaks in homonuclear 1H TOCSY or 1H NOESY spectra. By using the appropriate set of processing parameters we show that J
= 0.5(1/2) – MW/5000 + 1.8 for TOCSY spectra and J
= 0.6(1/2) – MW/5000 – 0.9 for NOESY spectra, where 1/2 is the half-height linewidth in Hz and MW is the molecular weight of the protein in Da. The simplicity of this relationship, combined with the ease with which 1/2 measurements can be made, means that J
coupling constants can now be rapidly determined (up to 100 measurements in less than 30 min) without the need for any complex curve-fitting algorithms. Tests on 11 different polypeptides involving more than 650 separate J
measurements have shown that this method yields coupling constants with an rmsd error (relative to X-ray data) of less than 0.9 Hz. Furthermore, the correlation coefficient between the predicted NMR coupling constants and those derived from high-resolution X-ray crystal structures is typically better than 0.89. These simple linear relationships have been found to be valid for peptides as small as 1 kDa to proteins as large as 20 kDa. Despite the method's simplicity, these results are comparable to the accuracy and precision of the best techniques published to date. 相似文献
64.
Hummel Herman Bogaards Roelof Bek Tatiana Polishchuk Lennard Amiard-Triquet Claude Bachelet Guy Desprez Michel Strelkov Peter Sukhotin Alex Naumov Andrei Dahle Salve Denisenko Stanislav Gantsevich Michael Sokolov Kirill Wolf Lein de 《Hydrobiologia》1997,355(1-3):127-138
The stress sensitivity, determined in copper exposureexperiments and in survival in air tests, and thegenetic structure, measured by means of isoenzymeelectrophoresis, were assessed in populations of theBaltic clam Macoma balthica (L.) from itssouthern to its northern distribution limit, in orderto test the hypotheses that near the distributionlimit the clams would be more stress sensitive andwould have a lower genetic variability. Thepopulations in west and north Europe show a stronggenetic resemblance. The populations in the sub-ArcticWhite Sea are genetically slightly different, and showa low stress sensitivity. The populations in theArctic Pechora Sea are genetically very distant fromthe other populations, and show the lowest stresssensitivity. Near the southern distribution limit, inagreement with the hypotheses, genetic variability islow and stress sensitivity high. On the other hand, incontrast to expectation, near the northerndistribution limit, in the populations of the PechoraSea, the genetic variability was higher, thus notreduced, and the stress sensitivity was low comparedto all other populations. Yet, it remains a questionif such is due to gradual physiologicalacclimatization (and ongoing differential selection)or to genetic adaptation. 相似文献
65.
Considerable controversy has surrounded the application of mitochondrial DNA data to reconstruction of evolutionary relationships
among the endemic cichlids of Lake Malawi. Central to this debate has been the issue of whether lineage sorting is complete,
and thus whether these data actually reflect species phylogeny, or simply gene genealogy. Review of all mtDNA control region
sequences available for members of one monophyletic subset of this species flock, the Malawi rockfishes, or mbuna, strongly indicates that lineage sorting is incomplete: Character-based analyses of these sequences reconstruct gene, not
species, interrelationships. Analysis of the pattern of nucleotide substitutions differentiating these mtDNA alleles suggests
that pyrimidine residues undergo transition substitutions more often than do purines. Estimation of the magnitude of derived
sequence differentiation in light of the reconstructed gene genealogy suggests that the mbuna may be of considerably more recent vintage than previous molecular characterizations have indicated.
Received: 6 April 1996 / Accepted: 3 March 1997 相似文献
66.
67.
68.
Dual nucleotide specificity of bovine glutamate dehydrogenase. The role of negative co-operativity.
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The thionicotinamide analogues of NAD+ and NADP+ were shown to be good alternative coenzymes for bovine glutamate dehydrogenase, with similar affinity and approx. 40% of the maximum velocity obtained with the natural coenzymes. Both thionicotinamide analogues show non-linear Lineweaver-Burk plots, which with the natural coenzymes have been attributed to negative co-operativity. Since the reduced thionicotinamide analogues have an isosbestic point at 340nm and have an absorption maximum at 400nm, it is possible to monitor reduction of natural coenzyme and thionicotinamide analogue simultaneously by dual-wavelength spectroscopy. When glutamate dehydrogenase is presented with NADP+ and thio-NADP+ simultaneously, the enzyme oligomer senses saturation of its coenzyme-binding sites irrespective of the exact nature of the coenzyme and locks the oligomer into its highly saturated form even when low saturation of the monitored coenzyme is present. These experiments substantiate the suggestion that glutamate dehydrogenase shows negative co-operativity in its catalytically active form. 相似文献
69.
Twenty isolates of Penicillium oxalicum produced secalonic acid as their major secondary metabolite. Fermentation conditions were determined for toxin production in grain and liquid media. The 50% lethal dose value for mice ranged from 26.5 to 51.7 mg/kg dependent on animal strain and sex, males being more susceptible than females. Secalonic acid was nontoxic and nonteratogenic to the chicken embryo and exhibited poor antibiotic properties. Its potential role in mycotoxicoses is discussed. 相似文献
70.