Degradation of the D1 protein of photosystem II under illumination in vivo: two different pathways involving cleavage or intermolecular cross-linking

The D1 protein of the photosystem II reaction center turns over the most rapidly of all the proteins of the thylakoid membrane under illumination in vivo. In vitro, the D1 protein sustained cleavage in a surface-exposed loop (DE loop) or cross-linking with another reaction center protein, the D2 protein or cytochrome b(559), under illumination. We found that the D1 protein was damaged in essentially the same way in vivo, although the resultant fragments and cross-linked adducts barely accumulated due to digestion by proteases. In vitro studies detected a novel stromal protease(s) that digested the adducts but not the monomeric D1 protein. These observations suggest that, in addition to cleavage, the cross-linking reactions themselves are processes involved in complete degradation of the D1 protein in vivo. Peptide mapping experiments located the cross-linking sites with the D2 protein among residues 226-244, which includes the cross-linking site with cytochrome b(559) [Barbato, R., et al. (1995) J. Biol. Chem. 270, 24032-24037], in the N-terminal part of the DE loop, while N-terminal amino acid sequencing of the fragment located the cleavage site around residue 260 in the C-terminal part of the loop. We propose a model explaining the occurrence of simultaneous cleavage and cross-linking and discuss the mechanisms of complete degradation of the D1 protein in vivo.     

Beta 3-adrenergic receptor is involved in feeding regulation in chicks

We examined whether the brain beta 3-adrenergic receptor (B3-AR) is involved in the feeding regulation of chicks. Intracerebroventricular (ICV) injection of BRL37344, a B3-AR agonist, reduced food intake of chicks under ad libitum, but not fasting, feeding conditions. The ICV injection of BRL37344 did not affect chick posture or locomotion activity suggesting that BRL37344 inhibited feeding without induction of sleep-like behavior as caused by norepinephrine. Furthermore, the rectal temperature increased following the ICV injection of BRL37344. Intraperitoneal administration of BRL37344 did not reduce food intake under ad libitum feeding condition. The present study demonstrated that the brain B3-AR is involved in the inhibition of feeding in chicks. We also suggested that activation of the brain affects the energy metabolism in chicks.     

The primary structure of the subunit in Bacillus thermoamyloliquefaciens KP1071 molecular weight 540,000 homohexameric alpha-glucosidase II belonging to the glycosyl hydrolase family 31

The gene that coded for the subunit of an molecular weight (Mr) 540,000 homohexameric alpha-glucosidase II (alpha-D-glucoside glucohydrolase, EC 3.2.1.20) produced by Bacillus thermoamyloliquefaciens KP1071 (FERM-P8477) growing at 30 to 66 degrees C was expressed in Escherichia coli HB101. The resulting homohexameric enzyme had a half-life of 10 min at 80 degrees C. Its purification and characterization showed that the enzyme was identical with the native one except for the latter deleting 7 N-terminal residues found in the former. The primary sequence of the subunit with 787 residues and an Mr of 91,070 deduced from the gene was 24-34% identical to the corresponding sequences of 15 alpha-glucosidases in the glycosyl hydrolase family 31 from 14 eukaryotic origins and the archaeon Sulfolobus solfataricus 98/2. From the sequence analysis by the neural network method of Rost and Sander [Rost, B. and Sander, C., Proteins: Struct. Funct. Genet., 19, 55-72 (1994)], we inferred that alpha-glucosidase II might make each subunit of 3 secondary structural regions, i.e., one N-terminal beta region, one central alpha/beta region with two catalytic residues Asp407 and Asp484, and one C-terminal beta region.     

Factors that affect the selection of tomato leaflets by two whiteflies,Trialeurodes vaporariorum and Bemisia tabaci (Homoptera: Aleyrodidae)

In Japan, although greenhouse whitefly, Trialeurodes vaporariorum (Westwood), and sweet potato whitefly, Bemisia tabaci (Gennadius), co-occur on tomato plants under greenhouse conditions, the two whiteflies are distributed differently with regard to leaf position. To elucidate the factors that determine the leaf position of these whiteflies, we investigated traits for leaflets collected from three positions on tomato plants. Furthermore, we examined leaflet selection by and fertility of the two whiteflies under choice and non-choice conditions. In addition, the effect on whitefly behavior of volatile compounds released from leaflets was evaluated by use of a Y-tube olfactometer test. Nitrogen and carbon content were highest for upper leaflets. In choice tests, more T. vaporariorum and B. tabaci adults selected upper and middle leaflets, respectively. Similarly, they oviposited more eggs on upper and middle leaflets. In non-choice tests, T. vaporariorum oviposited more eggs on upper leaflets, but B. tabaci oviposited equally on each leaflet. In Y-tube olfactometer tests, more T. vaporariorum adults moved to upper leaflets whereas more B. tabaci adults moved to middle leaflets. These results suggest that different leaflet selection by adults of these two whiteflies is likely to be associated with the different volatile compounds emitted by tomato leaflets at each position.     

Nano-sized manganese-calcium cluster in photosystem II

Cyanobacteria, algae, and plants are the manufacturers that release O₂ via water oxidation during photosynthesis. Since fossil resources are running out, researchers are now actively trying to use the natural catalytic center of water oxidation found in the photosystem II (PS II) reaction center of oxygenic photosynthetic organisms to synthesize a biomimetic supercatalyst for water oxidation. Success in this area of research will transcend the current bottleneck for the development of energy-conversion schemes based on sunlight. In this review, we go over the structure and function of the water-oxidizing complex (WOC) found in Nature by focusing on the recent advances made by the international research community dedicated to achieve the goal of artificial water splitting based on the WOC of PS II.     

Purification and characterization of a stable oxygen-evolving Photosystem II complex from a marine centric diatom, Chaetoceros gracilis

Oxygen-evolving Photosystem II particles (crude PSII) retaining a high oxygen-evolving activity have been prepared from a marine centric diatom, Chaetoceros gracilis (Nagao et al., 2007). The crude PSII, however, contained a large amount of fucoxanthin chlorophyll a/c-binding proteins (FCP). In this study, a purified PSII complex which was deprived of major components of FCP was isolated by one step of anion exchange chromatography from the crude PSII treated with Triton X-100. The purified PSII was still associated with the five extrinsic proteins of PsbO, PsbQ', PsbV, Psb31 and PsbU, and showed a high oxygen-evolving activity of 2135 μmol O₂ (mg Chl a)^− 1 h^− 1 in the presence of phenyl-p-benzoquinone which was virtually independent of the addition of CaCl₂. This activity is more than 2.5-fold higher than the activity of the crude PSII. The activity was completely inhibited by 3-(3,4)-dichlorophenyl-(1,1)-dimethylurea (DCMU). The purified PSII contained 42 molecules of Chl a, 2 molecules of diadinoxanthin and 2 molecules of Chl c on the basis of two molecules of pheophytin a, and showed typical absorption and fluorescence spectra similar to those of purified PSIIs from the other organisms. In this study, we also found that the crude PSII was significantly labile, as a significant inactivation of oxygen evolution, chlorophyll bleaching and degradation of PSII subunits were observed during incubation at 25 °C in the dark. In contrast, these inactivation, bleaching and degradation were scarcely detected in the purified PSII. Thus, we succeeded for the first time in preparation of a stable PSII from diatom cells.