Pancreatic lipase-colipase binds strongly to the thylakoid membrane surface.

BACKGROUND: Isolated thylakoid membranes, i.e. the photosynthetic membranes of green leaves, inhibit the activity of pancreatic lipase and colipase during hydrolysis of fat in vitro. This inhibition has been demonstrated to cause reduced food intake and improved hormonal and lipid profile in vivo. One of the reasons suggested for the inhibiting effect is binding of lipase-colipase to the thylakoid membrane surface. This prompted a study of the binding of lipase and colipase to thylakoids. RESULTS: The results showed that lipase and colipase strongly bind to the thylakoid membrane surface. The dissociation constant was determined at 1.2 × 10(-8) mol L(-1) ; binding decreased after treatment of thylakoids with pepsin/trypsin to 1.0 × 10(-7) and to 0.6 × 10(-7) mol L(-1) after treatment with pancreatic juice. Similarly, delipidation of thylakoids caused a decrease in binding, the dissociation constant being 2.0 × 10(-7) mol L(-1) . CONCLUSION: The binding of pancreatic lipase-colipase to the thylakoid membrane is strong and may explain the inhibition of lipase-colipase activity by thylakoids. After treatment with proteases to mimic intestinal digestion binding is decreased, but is still high enough to explain the observed metabolic effects of thylakoids in vivo. © 2013 Society of Chemical Industry

Violaxanthin accessibility and temperature dependency for de-epoxidation in spinach thylakoid membranes

Using DTT and iodoacetamide as a novel irreversible method to inhibit endogenous violaxanthin de-epoxidase, we found that violaxanthin could be converted into zeaxanthin from both sides of the thylakoid membrane provided that purified violaxanthin de-epoxidase was added. The maximum conversion was the same from both sides of the membrane. Temperature was found to have a strong influence both on the rate and degree of maximal violaxanthin to zeaxanthin conversion. Thus only 50% conversion of violaxanthin was detected at 4 degreesC, whereas at 25 degreesC and 37 degreesC the degree of conversion was 70% and 80%, respectively. These results were obtained with isolated thylakoids from non-cold acclimated leafs. Pigment analysis of sub-thylakoid membrane domains showed that violaxanthin was evenly distributed between stroma lamellae and grana partitions. This was in contrast to chlorophyll a and beta-carotene which were enriched in stroma lamellae fractions while chlorophyll b, lutein and neoxanthin were enriched in the grana membranes. In combination with added violaxanthin de-epoxidase we found almost the same degree of conversion of violaxanthin to zeaxanthin (73-78%) for different domains of the thylakoid membrane. We conclude that violaxanthin de-epoxidase converts violaxanthin in the lipid matrix and not at the proteins, that violaxanthin does not prefer one particular membrane region or one particular chlorophyll protein complex, and that the xanthophyll cycle pigments are oriented in a vertical manner in order to be accessible from both sides of the membrane when located in the lipid matrix

Supercapacitive Photo-Bioanodes and Biosolar Cells : A Novel Approach for Solar Energy Harnessing

The concept of supercapacitive photo-bioanode and biosolar cell (photo-biosupercapacitor) for simultaneous solar energy conversion and storage is demonstrated for the first time. Exploiting the capacitive component significantly improves the electron transfer processes and allows the achievement of a current density of 280 µA cm−2 in the pulse mode

Photoelectrochemical Communication between Thylakoid Membranes and Gold Electrodes through Different Quinone Derivatives

Photosynthesis is a sustainable process for the conversion of light energy into chemical energy. Thylakoids in energy-transducing photosynthetic membranes are unique in biological membranes because of their distinguished structure and composition. The quantum trapping efficiency of thylakoid membranes is appealing in photobioelectrochemical research. In this study, thylakoid membranes extracted from spinach are shown to communicate with a gold-nanoparticle-modified solid gold electrode (AuNP-Au) through a series of quinone derivatives. Among these, para-benzoquinone (PBQ) is found to be the best soluble electron-transfer mediator, generating the highest photocurrent of approximately 130 mu Acm(-2) from water oxidation under illumination. In addition, the photocurrent density is investigated as a function of applied potential, the effect of light intensity, quinone concentration, and amount of thylakoid membrane. Finally, the source of photocurrent is confirmed by using 3-(3,4-dichlorophenyl)-1,1-dimethylurea (known by its trade name, Diuron), an inhibitor of photosystem II, which decreases the total photocurrent by 50%

Pigments protect the light harvesting proteins of chloroplast thylakoid membranes against digestion by gastrointestinal proteases

Chloroplast thylakoid membranes inhibit pancreatic lipase/colipase activity in vitro and, when included in food, induce satiety signals. As thylakoid membranes themselves are nutrients, containing lipids and proteins, it is of interest to study the digestion of thylakoids by enzymes of the gastrointestinal tract. Thylakoid membranes were treated with pepsin, trypsin, gastric and pancreatic juice at 37 degrees C and the resulting enzymatic breakdown was analyzed by gel electrophoresis, electron microscopy and mass spectroscopy. In all cases, several of the proteins were degraded within half an hour, while the main parts of the pigment-protein complexes were resistant for hours. Oil emulsified thylakoids were more resistant towards the enzymatic breakdown. Electron microscopy demonstrated that, after treatments, the thylakoids still remained in a membrane vesicular form. The capacity of thylakoid membranes to inhibit the lipase/colipase activity was partly reduced in all cases. About 50% of the inhibition capacity remained after treatment with pancreatic juice when the thylakoids were present in an oil emulsion. Delipidated thylakoids and plasma membranes, which lack the photosynthetic pigments, were degraded rapidly by pancreatic juice. Conclusion: The pigments, closely bound to the trans-membrane helices of thylakoid membrane proteins protect these from digestion by pepsin, trypsin, gastric and pancreatic juice. This supports the notion that a substantial inhibition of lipase/colipase takes place during the first 2 h in the intestine resulting in a retardation and prolongation of lipolysis in vivo. (C) 2010 Elsevier Ltd. All rights reserved

Photocurrent Generation from Thylakoid Membranes on Osmium-Redox-Polymer-Modified Electrodes.

Thylakoid membranes (TMs) are uniquely suited for photosynthesis owing to their distinctive structure and composition. Substantial efforts have been directed towards use of isolated photosynthetic reaction centers (PRCs) for solar energy harvesting, however, few studies investigate the communication between whole TMs and electrode surfaces, due to their complex structure. Here we report on a promising approach to generate photosynthesis-derived bioelectricity upon illumination of TMs wired with an osmium-redox-polymer modified graphite electrode, and generate a photocurrent density of 42.4 μA cm(-2)