Conserved water-mediated H-bonding dynamics of catalytic Asn 175 in plant thiol protease

The role of invariant water molecules in the activity of plant cysteine protease is ubiquitous in nature. On analysing the 11 different Protein DataBank (PDB) structures of plant thiol proteases, the two invariant water molecules W1 and W2 (W220 and W222 in the template 1PPN structure) were observed to form H-bonds with the O_b atom of Asn 175. Extensive energy minimization and molecular dynamics simulation studies up to 2 ns on all the PDB and solvated structures clearly revealed the involvement of the H-bonding association of the two water molecules in fixing the orientation of the asparagine residue of the catalytic triad. From this study, it is suggested that H-bonding of the water molecule at the W1 invariant site better stabilizes the Asn residue at the active site of the catalytic triad.     

Na+ and K+ binding by glycerinated muscle fibers with reciprocal cation concentrations in the medium

The binding of Na+ and K+ by glycerinated muscle fibres was observed at reserve concentrations of NaCl in the medium. Under external concentrations of Na+ of K+ up to 0.4-0.5 mM, a constant fraction (0.15-0.25 mmoles/kg dry weight of the fibres) bound by glycerinated fibres was revealed. With the increase of NaCl or KCl concentration in the medium up to 10 mM the concentration of bound cations increased too. The parameters of Na+ and K+ sorption by glycerinated models were calculated. The values of Na+ and K+ binding limits were 4.4 and 1.8 mmole/kg dry weight of the fibres and those of affinity, 3.2 and 4.1 kcal/mol, respectively. The binding of one cation took place in conditions when its concentration was 10,000-20,000 fold less than that of the other cation. This points to the fact that Na+ and K+ binding is highly specific and is carried out by different centres. It is suggested that myosin ATPase is a substratum binding Na+ and K+ in glycerinated muscle fibres at reverse ratio concentrations of these cations in the medium.     

O-acetyl-salicylic acid promotes colony formation from protoplasts of an elite maize inbred

The salicylic acid derivative acetylsalicylic acid (ASA) was found to promote colony formation from protoplasts isolated from embryogenic suspension cultures of an elite maize inbred line. The drug was most effective at concentrations of 30–100 mg/l, and increases of more than 20-fold in the number of colonies recovered from protoplasts were obtained. The rate of growth of protoplast-derived cell colonies was not affected.Abbreviations 2,4-D 2,4-dichlorophenoxyacetic acid - DMSO dimethylsulfoxide - KM Kao and Michayluk medium (1975) - MES 2[N-morpholino]ethanesulfonic acid - ASA acetylsalicylic acid     

Interleukin inhibition by a parasite proteinase inhibitor, taeniaestatin

A proteinase inhibitor, taeniaestatin, isolated from the larval stage of the cestode Taenia taeniaeformis inhibits endogenous IL 2 generation in murine lymphocytes and IL 1 induced proliferation of murine thymocytes in a dose-dependent manner. However, taeniaestatin does not inhibit exogenous IL 2-induced proliferation of an IL 2-dependent cell line at any dose tested. These data indicate that the lack of IL 2 generation may be due in part to inhibition of a crucial cell-associated proteinase subsequent to cellular activation, or the lack of an effective IL 2 signal for differentiation. Our results are novel findings concerning molecular pathways for parasite inhibition of host immune responses, and suggest that selected proteinase inhibitors may be useful in clinical situations in which IL 1 or IL 2 are elevated.     

Chlorpromazine as permeabilizer and reagent for detection of microbial peroxidase and peroxidaselike activities

Chlorpromazine was used to perform a test for the detection of microbial peroxidase activities. The compound acts as both a cell permeabilizer and a reagent in the procedure developed which allows the detection of peroxidase and peroxidase like reactions both semiquantitatively in whole cell determinations and quantitatively in cell-free supernatants.     

Properties of aminoglycoside-3'-phosphotransferases determined by kanamycin transposones Tn 601 and Tn 5

Aminoglycoside-3'-phosphotransferase I and II (APT-3'-I and APT-3'-II) has been purified to homogenity from the cells of E. coli containing the plasmids R6 and JR67, respectively. The purification procedure involved competitive affinity chromatography on neomycin-sepharose and gel-filtration on Sephadex G-100. The specific activity of APT-3'-I with the substrates--lividomycin A, neomycin B, paromycin, ribostamycin, kanamycins A and B--are 4.3, 2.8, 2.1, 1.6, 0.9 and 0.8 mol/min. mg protein, respectively. The specific activity of APT-3'-II with the substrates--ribostamycin, paromycin, kanamycins A and B, neomycin B--are 8.0, 7.2, 4.0, 4.5 and 3.6, respectively. Mg2+ is required for the activity of both enzymes. Co2+, Zn2+ and Mn2+ are active in case of APT-3'-I; however, these cations are less active than Mg2+. The pH-optimum of APT-3'-I and APT-3'-II is 7.0--7.5. High ionic strength is required for the activity of both enzymes. The molecular weights of APT-3'-I and APT-3'-II are about 36 000 and 26 000, respectively. The amino acid composition of APT-3'-I and APT-3'-II was determined. Both enzymes contain tryptophane residues whose fluorescence intensity decreased when ATP, but not amino-glycoside antibiotics, is added. The interrelationship between the molecular weights of these enzymes and the sizes of the loops of transposones Tn 601 and Tn 5, encoding APT-3'-I and APT-3'-II, is discussed.     

X-ray absorption spectroscopy of selenium-containing amino acids

Received: 2 April 1999 / Accepted: 17 September 1999     

Lipid composition of spinal bone tissue in rabbits of different ages

Studies have been made on the lipid composition of total lipids, triglycerides and their fatty acids, cholesterol and phospholipids in the vertebral column of young and adult rabbits. It was shown that the content of total lipids and triglycerides increases, whereas that of cholesterol and phospholipids decreases with age. The content of total lipids in the vertebral column is 10 times higher as compared to that in the bones of the extremities. Mid-thoracic part of the vertebral column exhibits higher lipid content than other thoracic parts of the column. Lipid content of the vertebral processes is lower than that of the vertebral bodies. These data indicate lipid specificity and heterogeneity of bone tissue of the vertebral column. The main fatty acids of vertebral triglycerides are presented by those with 14-18 carbon atoms (90%), no acids with 22 atoms were found. Higher content of the linoleic acid (19%) and higher total unsaturation of triglycerides were found in the bone tissue of rabbits in comparison with those of man.