Protection and deprotection of horse cytochrome c

The last step in the semisynthesis of horse cytochrome c analogues (formation of the bond 65-66) requires the conformation of the complex between two complementary fragments, (1-65) lactone and (66-104). The fragments can be obtained from a limited degradation with cyanogen bromide. The amino component in this reaction can also be obtained from organo chemical synthesis in which the C-terminal fragment (81-104) is required in a selectively protected form. The latter is available from a cyanogen bromide degradation of ubiquitously protected cytochrome c. The details of the protection/deprotection reaction and the properties of nonadecamethylsulfonylethyloxycarbonyl cytochrome c are described.     

Verapamil counteracts the masking of long-lasting potentiation of hippocampal population spike produced by 2-amino-5-phosphonovalerate

In transversely sectioned rat hippocampal slices the effects of 2-amino-5-phosphonovalerate (APV), presumably a selective N-methyl-DL-aspartate antagonist, were examined on the development of long-lasting potentiation (LLP) of the CA1 population spike produced by Schaffer collateral tetanization (100 Hz, 1 s). APF (100 nA, 5 min), when applied 150-200 micron away from the CA1 cell bodies (apical dendritic area), had no significant effect if the population spike was evoked at 0.1 Hz, but produced a depression of the population spike if a 100 Hz tetanus was given during its iontophoresis. This depressant effect of APV was counteracted by verapamil (0.33 microM, 3 min) and LLP induced by the 100 Hz tetanus was unmasked. It is suggested that APV does not antagonize LLP, but only masks it by allowing CA++ influx into CA1 neurones that induces a depression. The results also raise doubts as to the selectivity of APV as an amino acid antagonist.     

Transfer of exogenous cholesterol to microsomes of hepatocytes investigated with [3H]desmosterol tracer.

The molybdenum centre of spinach (Spinacia oleracea) nitrate reductase has been investigated by e.p.r. spectroscopy of molybdenum(V) in reduced forms of the enzyme. The resting enzyme gives no signals attributable to Mo(V). However, on reduction with NADH, Mo(V) signals appeared at relatively short reaction times but decreased again on prolonged exposure to excess of the substrate as the enzyme was further reduced. On brief treatment of such samples with nitrate, Mo(V) signals reappeared but disappeared again on longer exposure to excess nitrate as the enzyme became fully reoxidized. Detailed investigation of the signals carried out in both 1H2O and 2H2O revealed the presence of two signal-giving species, referred to as 'signal A' and 'signal B', analogous to corresponding signals from nitrate reductase from Escherichia coli and from liver sulphite oxidase. Signal A has gav. 1.9767 and shows coupling to a single proton, exchangeable with the solvent, with A(1H)av. 1.3mT, whereas signal B shows no more than weak coupling to protons. Investigation of interconversion between the two species indicated that decreasing the pH from 8.0 to 6.7 had little effect, but that signal A was favoured by the presence of Cl-. This suggests, by analogy with recent work on sulphite oxidase by Bray, Gutteridge, Lamy & Wilkinson [Biochem. J. (1983) 211, 227-236] that Cl- is a ligand of molybdenum in the species giving signal A.     

Effect of ozone on ribosomes in pinto bean leaves

A study was made of cytoplasmic and chloroplast ribosomes in the primary leaves of pinto bean plants exposed to ozone. The isolated ribosomes were analysed by sucrose density gradient. Ozone at the levels of 0·35 ppm for 20–35 min does not change the concentrations of various sedimenting particles of the cytoplasmic ribosomes. Ozone at similar levels, however, specifically decreases the population of chloroplast ribosomes per unit fresh weight of leaves. The distribution pattern of these chloroplast ribosomes is characterized by the low concentration of the fast-sedimenting polysome particles concomitant with the low magnitude of other slow-sedimenting components. The kinetics of ribosome populations during leaf growth demonstrates that ozone does not influence the daily levels of different ribosomal components of cytoplasmic ribosomes. However, ozone prematurely decreases the concentrations of polysomes and other components of chloroplast ribosomes below control level at the early stage of leaf development. These findings are discussed to explain initiation of the premature senescence caused by ozone.     

Causes of Hypertension in the Young

Of 127 hypertensive patients aged 12 to 40 investigated by intravenous pyelography, abdominal aortography, and renal biopsy an underlying cause was found in 57%. The proportion with secondary hypertension was higher in young patients and in those with severe hypertension. Primary arteritis of the aorta was an important cause of renovascular hypertension in an Asian population.     

Single-step purification of pertussis toxin and its subunits by heat-treated fetuin-sepharose affinity chromatography

A general procedure for purifying biologically active pertussis toxin from Bordetella pertussis fermentation broth using affinity chromatography on heat-treated fetuin-Sepharose CL-4B is described. Diethanolamine is used as eluent in this single-step purification to prepare endotoxin-free pertussis toxin in good yield (70%) and high purity (greater than 95%). This one-step affinity chromatography procedure can be easily applied for large-scale preparation of pertussis toxin S1 subunit and its B-component. The affinity-purified S1 subunit is devoid of any of the histamine-sensitizing activity normally associated with pertussis toxin. The chromatographically purified pertussis toxin and its subunits retained their immunogenicity and could induce high levels of anti-toxin neutralizing antibodies.