Influence of structural and physical properties of the thylakoid membrane on QA - oxidation

Using isolated pea thylakoids, the relative rate of Q_A ^- oxidation has been estimated under various conditions, from the restoration of the induction curves following a dark period and from light 1-induced changes in modulated chlorophyll fluorescence excited by light 2.Alterations of _QinfA ^sup– oxidation rates were observed under conditions which affected the degree of thylakoid stacking, the lipid fluidity and the integrity of the membranes. The results are discussed in terms of the interactions between Q_A ^- and the plastoquinone pool with particular emphasis on lateral diffusion.Abbreviations DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea - EDTA Ethylenediaminetetracetate - Hepes N-2-hydroxyethyl piperazine-N-2-ethanesulphonic acid - NADP nicotinamide adenine dinucleotide phosphate     

Hormone and methylxanthine action on breast epithelial cells

Human mammary carcinoma cells and normal mouse breast epithelial cells desensitized as the result of treatment with beta-adrenergic agonists. Accumulation of cAMP in the same cells was affected only slightly by caffeine and there was no detectable desensitization or hypersensitization as a result of that treatment. However, as in many other cell types, caffeine was an effective inhibitor of adenosine action. These observations do not support the hypothesis made by Minton and his co-workers (1), that treatment of breast epithelial cells with agents that increase cAMP accumulation leads to hypersensitization rather than desensitization.     

Inhibition of protein carboxyl methylation by S-adenosyl-L-homocysteine in intact erythrocytes. Physiological consequences

S-Adenosyl-L-homocysteine was used to inhibit the methylation of carboxylic acid residues of membrane proteins in intact human erythrocytes. Incubation of erythrocytes for 24 h with 5 mM each of adenosine and L-homocysteine resulted in the intracellular accumulation of S-adenosyl-L-homocysteine and substantially inhibited membrane protein carboxyl methylation. From the degree of inhibition and from the observed turnover of methylated proteins, we estimate that the number of protein methyl esters in cells incubated with adenosine and L-homocysteine for 20 h is less than 20% that of cells incubated without these inhibitors. No significant differences in the physical deformability properties of the membrane of these hypomethylated cells were detected. However, there was a small but significant (p less than 0.001) increase in the amount of membrane protein D-aspartyl residues in these cells compared to control cells. These observations are consistent with the hypothesis that methylation of membrane proteins at D-aspartyl residues may result in the selective removal or repair of these uncommon residues.     

Studies of the mobility of maleimide spin labels within the erythrocyte membrane

We have confirmed a method yielding reproducible and reliable spectrometric parameters derived from spin-labeled erythrocyte ghosts using nitroxide derivatives of maleimide compounds. The disorder parameter, W/S, was shown to vary with changes in the structure of the label, the conditions utilized for labeling such as ionic strength and erythrocyte age and the presence of drugs such as alcohol and acetaminophen. The nitroxide spectrum was also found to change with increasing and decreasing temperature in an irreversible manner. These findings should permit increased reliance to be placed on the spin-labeling technique when used to monitor changes in membrane lipid or protein assembly.     

Development of the cytosolic defence system against microsomal lipid peroxidation in rat liver

Ascorbate-induced lipid peroxidation in rat liver microsomes reaches the adult level in 2-3 days. NADPH-induced peroxidation develops more gradually, in parallel with the activity of NADPH-cytochrome P-450 reductase, attaining adult levels by 10-12 days. The glutathione-dependent cytosolic enzyme activity which inhibits peroxidation is inhibited by bromosulphophthalein. The development of this system lags behind the development of microsomal lipid peroxidation between the ages of 2 and 20 days, allowing peroxidation to proceed.     

Lipid enrichment of thylakoid membranes. I. Using soybean phospholipids

(1) Using asolectin (mixed soybean phospholipids) liposomes, extra lipid, with or without additional plastoquinone, has been introduced into isolated thylakoid membranes of pea chloroplasts. (2) Evidence for this lipid enrichment was obtained from freeze-fracture which indicated that a decrease in the numbers of EF and PF particles per unit area of membrane occurred with increasing lipid incorporation. The decrease was not due to loss of integral membrane polypeptides as judged by assay of cytochrome present or SDS-polyacrylamide gel electrophoresis of lipid-enriched membrane fractions. Moreover, the enrichment procedure did not lead to extraction of low molecular weight lipophilic membrane components or of thylakoid membrane lipids. (3) The introduction of phospholipids into the membrane affected steady-state electron transport. Inhibition of electron transport was observed when either water (Photosystem (PS) II + PS I) or duroquinol (PS I) was used as electron donor with methyl viologen as electron acceptor, and the degree of inhibition increased with higher enrichment levels. Introduction of exogenous plastoquinone with the additional lipid had little effect on whole-chain electron transport, but caused an increase in the 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone (DBMIB)-sensitive rate of PS I electron transport. The inhibition was also detected by flash-induced oxidation-reduction changes of cytochrome f.     

The optical properties of CuA in bovine cytochrome c oxidase determined by low-temperature magnetic-circular-dichroism spectroscopy.

The visible-near-i.r.-region m.c.d. (magnetic-circular-dichroism) spectrum recorded at low temperature in the range 450-900 nm is reported for oxidized resting mammalian cytochrome c oxidase. M.c.d. magnetization curves determined at different wavelengths reveal the presence of two paramagnetic species. Curves at 576, 613 and 640 nm fit well to those expected for an x,y-polarized haem transition with g values of 3.03, 2.21 and 1.45, i.e. cytochrome a3+. The m.c.d. features at 515, 785 and 817 nm magnetize as a S = 1/2 paramagnet with average g values close to 2, and simulated m.c.d. magnetization curves obtained by using the observed g values of CuA2+, i.e. 2.18, 2.03 and 1.99, fit well to the experimental observations. The form of the m.c.d. magnetization curve at 466 nm is curious, but it can be explained if CuA2+ and cytochrome a3+ contribute with oppositely signed bands at this wavelength. By comparing the m.c.d. spectrum of the enzyme with that of extracted haem a-bisimidazole complex it has been possible to deconvolute the m.c.d. spectrum of CuA2+, which shows transitions throughout the spectral region from 450 to 950 nm. The m.c.d.-spectral properties of CuA2+ were compared with those of a well-defined type I blue copper centre in azurin isolated from Pseudomonas aeruginosa. The absolute intensities of the m.c.d. signals at equal fields and temperatures for CuA2+ are 10-20-fold greater than those for azurin. The optical spectrum of CuA2+ strongly suggests an assignment as a d9 ion rather than Cu(I) bound to a thiyl radical.     

EVIDENCE FOR A CRYPTOMONAD SYMBIONT IN THE CILIATE,CYCLOTRICHIUM MEUNIERI1

Extracts of the marine ciliate Cyclotrichium meunieri contained chlorophylls a and c, carotenoids, and a phycoerythrin with a single absorbance maximum at 542 nm. This assemblage of pigments suggests that the numerous photosynthetic symbionts present in each ciliate cell belong to the Cryptophyceae.     

The fine structure of the sense organs of the cephalopod mollusc Nautilus

Summary The structure of the rhinophore, digital tentacles, post-ocular tentacles and the eye of Nautilus macromphalus are described. The rhinophore is composed of mucous cells, ciliated cells, and flask-shaped ciliated cells. The latter are probably olfactory receptors. The digital tentacles are composed of mucous cells and pigmented cells. Motor-end-plates found in the muscle layer below the epithelium of the digital tentacles are similar to those described in other cephalopods. The post-ocular tentacle contains receptor cells that bear macrocilia. These may be mechanoreceptors. The retina is composed of retinula cells and supporting cells. A complex rhabdom is formed at the distal ends of the retinula cells. The supporting cells send processes up between these rhabdoms. Both types of cells contain pigment granules but the retinula cell has a complex membranous structure in its perikaryon. No synapses were found at the bases of the retinula cells. At the side of the retina are mucous cells that are presumed to produce the jelly-like substance that fills the inside of the eye in life. The likely function of the eye is discussed and it is suggested that it is capable of simple discriminations. It is suggested that the sense organs are probably comparatively unchanged from those of fossil nautiloids. Acknowledgements. This paper is dedicated to the late Dr. Yves Merlet who collected the nautiluses used in this study.We would like to thank Prof. J. Z. Young for all his support and encouragement. The Royal Society, The Percy Sladen Memorial Fund, and University College, London, provided the financial support that enabled one of us (V.C.B.) to collect nautiluses. The Science Research Council, U.K., provided the electron microscope used in the major part of the study and a grant to one of us (V.C.B.). We would also like to thank Prof. J. B. Gilpin-Brown who provided Fig. 1, Dr. R. Catala, for aquarium facilities, Mr. M. P. Legand and the Institut Français d'Oceanie, Noumea, New Caledonia, for laboratory facilities, Dr. J.-M. Bassot and Dr. Anna Bidder for advice on catching and preserving nautiluses, Mrs. Judy Parkes and Mr. M. Barker for photographic assistance, and Miss J. Date for secretarial assistance.