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Tryptophan-scanning mutagenesis of the ligand binding pocket in Thermotoga maritima arginine-binding protein.
We constructed single tryptophan mutants to probe the arginine binding pocket. Trp residues placed around the binding pocket reveal a strong dependence on fluorescence emission of the protein with arginine for all but one of the mutants. Using these data, we calculated dissociation constants of 1.9-...
Cathepsin D activity and selectivity in the acidic conditions of a tumor microenvironment: Utilization in the development of a novel Catheps...
We investigated the effect of pH on the catalytic activity and selectivity of mature Cathepsin D (CD). It was found that the increase in the pH of the media led to a decrease in the reaction rate. However, the specificity of mature CD was not affected by a variation in pH. In the second step, the ef...
Site directed mutagenesis of StSUT1 reveals target amino acids of regulation and stability.
We provide further experimental evidence for a pH optimum at physiological extrema. Site directed mutagenesis provides information about functional amino acids, which are highly conserved and responsible for this extraordinary increase in transport capacity under extreme pH conditions. Redox-depende...
The eubacterial protein synthesis inhibitor pulvomycin interacts with archaeal elongation factor 1α from Sulfolobus solfataricus.
The effect of pulvomycin on the biochemical and fluorescence spectroscopic properties of the archaeal elongation factor 1α from Sulfolobus solfataricus (SsEF-1α), the functional analog of eubacterial EF-Tu, was investigated. The antibiotic was able to reduce in vitro the rate of protein synthesis...
Mode of action of the antiprion drugs 6AP and GA on ribosome assisted protein folding.
We have characterized with biochemical experiments, the mode of inhibition of these two drugs using ribosomes or ribosomal components active in protein folding (referred to as 'ribosomal folding modulators' or RFMs) from both bacteria Escherichia coli and yeast Saccharomyces cerevisiae, and human ca...
A chymotrypsin-like proteinase from the midgut of Tenebrio molitor larvae.
A chymotrypsin-like proteinase was isolated from the posterior midgut of larvae of the yellow mealworm, Tenebrio molitor, by ion-exchange and gel filtration chromatography. The enzyme, TmC1, was purified to homogeneity as determined by SDS-PAGE and postelectrophoretic activity detection. TmC1 had a...
Expression of eukaryotic glycosyltransferases in the yeast Pichia pastoris.
We have examined the expression in P. pastoris of Arabidopsis thaliana and Drosophila melanogaster core alpha1,3-fucosyltransferases (EC 184.108.40.206), A. thaliana beta1,2-xylosyltransferase (EC 220.127.116.11), bovine beta1,4-galactosyltransferase I (EC 18.104.22.168), D. melanogaster peptide O-xylosyltransferas...
The human TINF2 gene organisation and chromosomal localization.
We need to understand how the genes that control telomere length are regulated. TIN2, the protein product of the human TINF2 gene, was recently identified as a negative regulator of telomere length. Here I report the organisation and chromosomal localization of the TINF2 gene. It contains six exons...
Expression of the Daucus carota somatic embryogenesis receptor kinase (DcSERK) protein in insect cells.
The Daucus carota somatic embryogenesis receptor kinase (DcSERK) gene serves as marker to monitor the transition from somatic into embryogenic plant cells. To determine the intrinsic biochemical properties of the DcSERK protein, a predicted transmembrane receptor, the kinase domain was expressed as...
Colloidal aggregates of insoluble inclusions in human goiters.
We have screened retrospectively the autoradiographs of 60 human nodular goiters labeled 17 h preoperatively with 100 microCi 125I for evidence of colloid compartmentalization. In 87% (52/60) of all goiters examined, sporadic or multiple colloidal inclusions ('colloid stones') not mixing with newly...