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In the present study we have analysed the potential relationship between the cellular level of adenine nucleotides and the activity of microsomal cholesterol ester hydrolase by treating rat hepatocyte suspensions with fructose or adenosine. Fructose raised the microsomal hydrolysis of cholesteryl esters as a function of the dose. This ketose led to marked decreases in the cell level of ADP, ATP...
Oligodeoxynucleotides (ONs) are currently being tested in clinical trials as anti-viral and thrombolytic agents. Although ONs are biologically active in almost every cell line examined, these molecules are inefficiently internalized. In addition, the mechanism of ON transport to the active site in either the cytoplasm or nucleus remains ill-defined. Thus, elucidation of the mechanism of ON cellular...
A convenient method to purify each of the non-ribosomal proteins required to translate a native mRNA in vitro is described. In this scheme, the ribosome is used as an 'affinity' matrix to selectively elute the non-ribosomal proteins required for translation that are bound to these particles. Different sets of these proteins can be eluted with solutions of Mg 2+ and NH +4 ...
Reactions of hydroxyl radicals with horseradish peroxidase (HRP) have been studied by means of pulse radiolysis technique in the absence and presence of oxygen. Hydroxyl radicals, produced in excess towards enzyme, react exclusively with the protein part of HRP with the rate constant k = 1.1 x 10 11 M -1 s -1 . Activity loss induced by OH * is connected...
The Lactobacillus leichmannii pyrB gene, encoding pyrimidine biosynthetic enzyme aspartate transcarbamylase (ATCase), was cloned from a partial genomic library lying on a 1468 bp SalI/BstXI fragment. The predicted polypeptide sequence extending over 351 amino acid residues (M r 39 855 Da) was compared to those of various other organisms revealing clear identities towards them and important...
Histone H4 stimulates the uptake of glucose in rat adipocytes and muscle cells. However, the mechanism of this unusual activity is not known. Therefore, we have begun to investigate the mechanism by which histone H4 stimulates the glucose uptake in rat adipocytes. We report that histone H4 requires 15-20 min to achieve its maximum effect and its time course is virtually indistinguishable from the...
Acetylcholinesterase (AChE) has been identified and purified from the venom of desert cobra (W aegyptia) to apparent homogeneity using a TSK G 3000 SW gel filtration column and a Mono Q anion-exchange column. AChE was purified to homogeneity as established by sodium dodecylsulfate/polyacrylamide gel electrophoresis. The specific activity of AChE was 357 IU/mg with acetylthiocholine iodide as substrate...
Recombinant vasoactive intestinal polypeptide (VIP) analogs were expressed in Escherichia coli as a fusion protein containing tandemly repeated multiple copies of a synthetic VIP gene joined to glutathione S-transferase. The encoded protein contains VIP units separated by a linker peptide, potentially excisable by a double cleavage with endoprotease factor Xa and hydroxylamine. Expression of different...
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