Identification of biochemical association of phycobilisome with photosystems in cyanobacterial state transition

State transition is a short-term balance mechanism of energy distribution between photosystem II （PSII） and PSI. Although light-induced state transition in cyanobac- teria has been suggested to depend completely on the phy- cobilisome （PBS） movement between PSII and PSI, the biochemical evidence has not been clearly shown. In this study, we locked the association of PBS with PSII or PSI using glycinebetaine when cells attain State 1 or 2 by expos- ure to light of blue or green, respectively. Subsequently, the PBS-reaction centers were resolved by blue native poly- acrylamide gel electrophoresis and two-dimensional elec- trophoresis, and then identified by western blot analysis. The results showed that in wild-type （WT） Synechocystis sp. strain PCC 6803, the PBS core always co-migrates with the PSII dimer during light-induced State 1-State 2 transi- tion, but its rod leaves the PSII dimer in State 2 regardless of its co-migration in State 1. In the light-induced State 2, the co-migration of PBS rod with PSI trimer was observed in WT, but not in AndhB （M55）, a State-2-transition-defi- cient mutant. This study first provided the biochemical evi- dence for the association of PBS with photosystems during cyanobacterial state transition.     

Properties of serine：glyoxylate aminotransferase purified from Arabidopsis thaliana leaves

The photorespiratory enzyme L-serine：glyoxylate amino- transferase （SGAT; EC 2.6.1.45） was purified from Arabidopsis thaliana leaves. The f＇mal enzyme was approximately 80 % pure as revealed by sodium dodecyl sulfatepolyacrylamide gel electrophoresis with silver staining. The identity of the enzyme was confirmed by LC/MS/MS analysis. The molecular mass estimated by gel filtration chromato- graphy on Sephadex G-150 under non-denaturing conditions, mass spectrometry （matrix-assisted laser desorption/ ionization/time of flight technique） and sodium dodecyl sulfate-polyacrylamide gel electrophoresis was 82.4 kDa, 42.0 kDa, and 39.8 kDa, respectively, indicating dimer as the active form. The optimum pH value was 9.2. The enzyme activity was inhibited by aminooxyacetate and β-chloro-L-alanine both compounds reacting with the carbonyl group of pyridoxal phosphate. The enzyme＇s transaminating activity with L-alanine and glyoxylate as substrates was approximately 55 % of that observed with L-serine and glyoxylate. The lower Kmvalue （1.25 mM） for L-alanine, compared with that of other plant SGATs, and the kcat/Km（Ala） ratio being approxi- mately 2-fold higher than kcat/Km（Ser） suggested that, during photorespiration, Ala and Ser are used by Arabidopsis SGAT with equal efficiency as amino group donors for glyoxylate. The equilibrium constant （Keq）, derived from the Haldane relation, for the transamination reaction between L-serine and glyoxylate with the formation of hydroxypyruvate and glycine was 79.1, strongly favoring glycine synthesis. However, it was accompanied by a low Km value of 2.83 mM for glycine. A comparison of some kinetic properties of the studied enzymes with the recombinant Arabidopsis SGATs previously obtained revealed substantial differences. The ratio of the velocity of the transamination reaction with L-alanine and glyoxylate as substrates versus that with L-serine and glyoxylate was 1：1.8 for the native enzyme, whereas it was 1：7 for the recombinant SGAT. Native SGAT showed a much lower Km value for L-alanine compared to the recombinant enzyme.     

Refined structure of basic phospholipase A2 from venom of Agkistrodon halys Pallas in orthorhombic crystal form I at 0.25 nm resolution

The basic phospholipase A2 from the venom of Agkistrodon halys Pallas is a potent hemolytic toxin and anticoagulant. The accurate rotation and translation parameters of the molecules in orthorhombic crystal form I were successfully obtained using the fitting refinement technique. The structure was refined in the resolution range of 0. 6-0.25 nm using least square refinement with non-crystallographic two fold symmetry restraint, and resulted in the final R factor of 20.1 % , and the rms deviations from ideal stereochemistry were 0. 001 3 nm for bond lengths and 1. 32° for bond angles. The overall architecture of the present structure was similar to that of the determined structure of the orthorhombic crystal form Ⅱ, with a few differences in the regions of the β-wing and Ca²⁺-binding loop. The dimers formed by the two molecules in the asymmetric unit in both crystal forms were also similar. However, one of the monomers showed an orientational difference of 5.5° along the dimer interface in the two crystal forms, suggesting the flexibility of the interface of the dimer to some degree. The molecular packing of the dimer in crystal form I was much more compact than that in crystal form Ⅱ.     

Crystal structure of Cas1 in complex with branched DNA

Cas1 is a key component of the CRISPR adaptation complex, which captures and integrates foreign DNA into the CRISPR array,resulting in the generation of new spacers. We have determined crystal structures of Thermus thermophilus Cas1 involved in new spacer acquisition both in complex with branched DNA and in the free state. Cas1 forms an asymmetric dimer without DNA.Conversely, two asymmetrical dimers bound to two branched DNAs result in the formation of a DNA-mediated tetramer, dimer of structurally asymmetrical dimers, in which the two subunits markedly present different conformations. In the DNA binding complex, the N-terminal domain adopts different orientations with respect to the C-terminal domain in the two monomers that form the dimer. Substrate binding triggers a conformational change in the loop 164–177 segment. This loop is also involved in the 3′ fork arm and 5′ fork arm strand recognition in monomer A and B, respectively. This study provides important insights into the molecular mechanism of new spacer adaptation.     

苦瓜降糖成分研究论文被《自然中国》列为研究亮点

上海药物研究所天然药化室叶阳课题组和澳大利亚伽尔文（Garvan）医学研究所合作的研究论文Antidiabetic activities of triterpenoids isolated from bitter melon associated with activation of the AMPK pathway（Tan,M.J.et al.,Chem.Biol.2008,15,263-73）被《自然中国》列为研究亮点。苦瓜是日常食用的蔬菜,明代《滇南本草》就记载称苦瓜有“清暑、益气、止渴”的功效。     

The Quaternary Structure of NADPH Thioredoxin Reductase C Is Redox-Sensitive

NADPH thioredoxin reductase C （NTRC） is a chloroplast enzyme able to conjugate NADPH thioredoxin reductase （NTR） and thioredoxin （TRX） activities for the efficient reduction of 2-Cys peroxiredoxin （2-Cys PRX）. Because NADPH can be produced in chloroplasts during darkness, NTRC plays a key role for plant peroxide detoxification during the night. Here, it is shown that the quaternary structure of NTRC is highly dependent on its redox status. In vitro, most of the enzyme adopted an oligomeric state that disaggregated in dimers upon addition of NADPH, NADH, or DTT. Gel filtration and Western blot analysis of protein extracts from Arabidopsis chloroplast stroma showed that native NTRC forms aggregates, which are sensitive to NADPH and DTT, suggesting that the aggregation state might be a significant aspect of NTRC activity in vivo. Moreover, the enzyme is localized in clusters in Arabidopsis chloroplasts. NTRC triple and double mutants, A164G- V182E-R183F and A164G-R183F, replacing key residues of NADPH binding site, showed reduced activity but were still able to dimerize though with an increase in intermediary forms. Based on these results, we propose that the catalytically active form of NTRC is the dimer, which formation is induced by NADPH.     

Substrate Selectivity of Glycerol-3-phosphate Acyl Transferase in Rice

Substrate selectivity of glycerol-3-phosphate acyltransferase （EC 2. 3. 1. 15） of rice （Oryza sativa L.） was explored in a comparative study of acyltransferases from seven plant species. In vitro labeling of acyl carrier protein （ACP） with ^14C or 3H showed that acyltransferase from chill-sensitive plants, such as rice that uses either oleic （18：1） or palmitic acid （16：0） as acyl donor at comparable rates, displays lower selectivity than the enzyme from chill-resistant plants, such as spinach, which preferentially uses oleic acid （18：1） rather than palmitic acid （16：0） as an acyl donor. This may be a result of the size and character of the substrate-binding pocket of acyltransferase. Homology modeling and protein structure-based sequence alignment of acyltransferases revealed that proteins from either chill-sensitive or chill-tolerant plants shared a highly conserved domain containing the proposed substrate-binding pocket. However, the aligned residues surrounding the substrate-binding pocket are highly heterogeneous and may have an influence mainly on the size of the substrate binding pockets of acyltransferases. The substrate selectivity of acyltransferase of rice can be improved by enlarging the substrate-binding pocket using molecular biological methods.     

Structure of desheptapeptide (B24-B30) insulin in a new crystal form

The structure of desheptapeptide (B24-B30) insulin (DHPI) in a new crystal form (form B) has been determined and refined to 0.2 nm resolution. The crystals were obtained under the same crystallization condition as previously reported crystal form (form A). The overall structures of the two crystal forms are similar but obvious differences can be observed in crystal packing and local conformation. The crystal structures of the two forms show that the two independent molecules in an asymmetric unit from a DHPI dimer, and the dimer formation buries more than 18.20 and 16.95 nm~2 of solvent accessible surfaces for form A and form B DHPI, respectively, the largest among insulin and insulin analogs ever reported. Close examination at crystal packing shows that the dimer-forming surface of DHPI, namely Surface Ⅱ, is normally present in the association of insulin and insulin analogs in their crystal structures. The results demonstrate that Surface Ⅱ is crucially important for the formation of two crystal form     

Structural Basis of Interaction of Bipartite Nuclear Localization Signal from Agrobacterium VirD2 with Rice Importin-o

Dear Editor, In the eukaryotic cell, the nuclear envelope separates genomic DNA from the rest of the cell, but ensures a molecu- lar communication between the nucleus and the cytoplasm through the nuclear pore complexes （NPCs）. Nuclear trans- port of macromolecules is an energy-dependent process, exemplified by the classical nuclear import pathway. In the cytoplasm, proteins containing classical nuclear localization signals （cNLSs） are recognized by a hetero-dimeric transport receptor （importin-α （Impα）：importin-β （Impl3） complex）. Impa is employed as an adaptor protein that specifically rec-ognizes the classical nuclear localization signal （cNLS） from the cargo protein （Marfori et al., 2011）. Translocation of the trimeric cargo：transport receptor import complex through the NPCs is mediated by transient interactions between Impβ and nucleoporins in the NPC. The directionality of the trans- port is imparted by the asymmetric distribution, between the cytoplasm and the nucleus, of the distinct nucleotide-bound （GTP/GDP） states of the small GTPase Ran （Tran et al., 2007）.     

PNAS：单细胞测序技术应用于癌症无创诊断

2013年12月10日著名学术期刊美国《国家科学院刊）（Proceedings of the National Academy of SciencesUSA）在线发表了北京大学生命科学学院生物动态光学成像中心谢晓亮、白凡课题组与北大肿瘤医院王洁团队合作的研究结果。     

Encounters with aphid predators or their residues impede searching and oviposition by the aphid parasitoid Aphidius ervi （Hymenoptera： Aphidiinae）

Intraguild predation （IGP） can be an important factor influencing the effective- ness of aphid natural enemies in biological control. In particular, aphid parasitoid foraging could be influenced by the presence of predators. This study investigated the effect of larvae of the predatory hoverfly Episyrphus balteatus DeGeer （Diptera： Syrphidae） and the multicolored Asian ladybird Harmonia axyridis Pallas （Coleoptera： Coccinellidae） on the foraging behavior of the aphid parasitoid, Aphidius ervi Haliday （Hymenoptera： Aphidiidae） in choice experiments using a leaf disc bioassay. Wasp response to chemical tracks left by those predator larvae was also tested. Parasitoid behavior was recorded using the Observer （Noldus Information Technology, version 5.0, Wageningen, the Netherlands）. The experiments were conducted under controlled environmental conditions using leaves of the broad bean plant, Viciafaba L. （Fabaceae） with Myzus persicae Sulzer （Homoptera： Aphididae） as the host complex. A. ervi females avoided aphid patches when larvae of either predator were present. A similar avoidance response was shown by A. ervi to aphid patches with E. balteatus larval tracks, whereas no significant response was observed to tracks left by H. axyridis larvae. It was concluded that IG predator avoidance shown by the aphid parasitoid A. ervi may be a factor affecting their distribution among host patches.     

Effects of Cadmium on Polar Lipid Composition and Unsaturation in Maize （Zea mays） in Hydroponic Culture

This study aimed to evaluate the effect of Cd exposure （100 μmol/L） on polar lipid composition, and to examine the level of fatty acid unsaturation in maize （Zea mays L.）. In roots, the level of 16：0 and monounsaturated fatty acids （16：1 ＋ 18：1） decreased in phosphatidylcholine （PC） and phosphatidylethanolamine （PE）. In contrast, the proportion of unsaturated 18-C fatty acid species showed an opposite response to Cd. The content, on the other hand, of PC, PE, digalactosyldiacylglycerol （DGDG）, and steryl lipids increased in roots （2.9-, 1.6-, 5.3-, and 1.7-fold increase, respectively）. These results suggest that a more unsaturated fatty acid composition than found in control plants with a concomitant increase in polar lipids may favor seedling growth during Cd exposure. However, the observed increase in the steryl lipid （SL） ： phospholipid （PL） ratio （twofold）, the decrease in monogalactosyldiacylglycerol （MGDG） ： DGDG ratio, as well as the induction of lipid peroxidation in roots may represent symptoms of membrane injury. In shoots, the unsaturation level was markedly decreased in PC and phosphatidylglycerol （PG） after Cd exposure, but showed a significant increase in sulfoquinovosyldiacylglycerol （SQDG）, MGDG and DGDG. The content of PG and MGDG was decreased by about 65%, while PC accumulated to higher levels （4.4-fold increase）. Taken together, these changes in the polar lipid unsaturation and composition are likely to be due to alterations in the glycerolipid pathway. These results also support the idea that the increase in overall unsaturation plays some role in enabling the plant to withstand the metal exposure.     

新型雄激素受体调节剂研究论文被《自然中国》列为研究亮点

2008年5月7日,依托于上海药物研究所的国家新药筛选中心王明伟课题组和四川大学华西药学院合作的研究论文“Discovery and biological characterization of a novel series of androgen receptor modulators”（Zhou,C.et al.,Bri.J.Pharmcol.,doi：10.1038/bjp.2008.107）被《自然中国》列为“研究亮点”：一类新型雄激素受体调节剂或能更好地治疗前列腺癌。     

Introduction to Asian Herpetological Research

Asian Herpetological Research （AHR）, an international English language journal, is published quarterly by the Chengdu Institute of Biology （CIB）, Chinese Academy of Sciences and the Science Press of China, cooperated with the Asiatic Herpetological Research Society （AHRS）, with its registered numbers： CN 51-1735/Q and ISSN 2095-0357, and post distribution code： 62-218. AHR has an international Editorial Board consisting of many top herpetologists from different countries in the world. The journal＇s website can be found at： http：//www.ahr-journal.com.     

Introduction to Asian Herpetological Research

Asian Herpetological Research （AHR）, an international English language journal, is published quarterly by the Chengdu Institute of Biology （CIB）, Chinese Academy of Sciences and the Science Press of China, cooperated with the Asiatic Herpetological Research Society （AHRS）, with its registered numbers： CN 51-1735/Q and ISSN 2095-0357, and post distribution code： 62-218. AHR has an international Editorial Board consisting of many top herpetologists from different countries in the world. The journal＇s website can be found at： http：//www.ahr-journal.com.     

Nutrient content affecting Spodoptera frugiperda and Dalbulus maidis occurrence in corn

This work aims to study the effect of the nutrient contents in the leaves of corn on the incidence of Spodopterafrugiperda （Smith） （Lepidoptera： Noctuidae） and Dalbulus maidis （Delong ＆ Wolcott） （Homoptera： Cicadellidae）. The treatments were represented by unfertilized corn plants and corn plants fertilized with 250 kg of the formulation 4-14-8 （N- P-K） ＋ 100 kg of ammonium sulfate （AS）/ha and 500 kg of the formulation 4-14-8 （N-P-K） ＋200 kg of AS/ha, arranged in randomized blocks with four replicates. Attacks by S. frugiperda and D. maidis were higher as the nitrogen, sulfur, calcium and copper content in the leaves of corn increased. The effects of calcium and copper found in this study were not consistent with that of lower susceptibility associated with higher levels of these nutrients mentioned in the literature. We believe that there was a cumulative effect of the nutrients altogether, building up a nutritiously better vegetable which in turn had less reserve for defense, or that the content of both nutrients in the leaves were not high enough to induce such effects.     

Host plant mediation of diapause induction in the cabbage beetle, Colaphellus bowringi Baly （Coleoptera： Chrysomelidae）

The cabbage beetle, Colaphellus bowringiBaly （Coleoptera： Chrysomelidae）, is a serious pest of crucifers in China, undergoing an imaginal summer and winter diapause in the soil. The effects of host plants on diapause incidence were tested in the beetle. The ratio of adults entering diapause was significantly low when they fed on the mature leaves of Chinese cabbage Shanghaiqin （Brassica chinesis var communis） than those feeding on Chinese cabbage Suzhouqin （Brassica chinesis var communis）, radish （Raphanus sativus var longipinnatus） and stem mustard （Brassicajuncea vat tumida） at 25℃ combined with 13：11 （L： D） h. Fewer adults entered diapause on young leaves compared to physiologically aged and mature radish leaves at 25℃ combined with 13：11 （L： D） h. The effect of host plant species on diapause induction was also evident under continuously dark rearing conditions or at different photoperiods. These experimental results demonstrate that host plant mediation of diapause induction exists in the cabbage beetle. However, at temperatures ≤20℃ or photoperiods of 16：8 （L： D） h combined with 25℃, all individuals entered diapause regardless of the host plants, indicating that the effects of host plants on diapause induction could be expressed only within a limited range of temperatures and photoperiods.     

The integrated use of chemical insecticides and the entomopathogenic nematode,Steinemema carpocapsae （Nematoda： Steinernematidae）, for the control of sweetpotato whitefly,Bemisia tabaci （Hemiptera： Aleyrodidae）

The integration of chemical insecticides and infective juveniles of the entomopathogenic nematode Steinernema carpocapsae （Wesier） （Nematoda： Steinernematidae）, to control second instars of the sweetpotato whitefly, Bemisia tabaci Gennadius （Hemiptera： Aleyrodidae） was investigated. Using a sand bioassay, the effects of direct exposure of S. carpocapsae for 24 h to field rate dilutions of four insecticides （spiromesifen, thiacloprid, imidacloprid and pymetrozine） on infectivity to Galleria mellonella larvae were tested. Although all chemicals tested, except spiromesifen, produced acceptable nematode infectivity rates, they were all significantly less than the water control. The effect of insecticide treatment （dry residues of spiromesifen, thiacloprid and pymetrozine and soil drench of imidacloprid） on the efficacy of the nematode against B. tabaci was also investigated. Nematodes in combination with thiacloprid and spiromesifen gave higher B. tabaci mortality （86.5% and 94.3% respectively） compared to using nematodes alone （75.2%） on tomato plants. There was no significant difference in B. tabaci mortality when using the chemicals imidacloprid, pymetrozine and spiromesifen in conjunction with nematodes compared to using the chemicals alone. However, using thiacloprid in combination with the nematodes produced significantly higher B. tabaci mortality than using the chemical alone. The integration of S. carpocapsae and these chemical agents into current integrated pest management programmes for the control of B. tabaci is discussed.     

Introduction to Asian Herpetological Research

Asian Herpetological Research （AHR）, an international English language joumal, is published quarterly by the Chengdu Institute of Biology （CIB）, Chinese Academy of Sciences and the Science Press of China, cooperated with the Asiatic Herpetological Research Society （AHRS）, with its registered numbers： CN 51-1735/Q and I SSN 2095-0357, and post distribution code： 62-218. AHR has an intemational Editorial Board consisting of many top herpetologists from different countries in the world. The joumal＇s website can be found at： http：//www.ahr-joumal.com.