Antifreeze glycopeptide adsorption on single crystal ice surfaces using ellipsometry

Antarctic fishes synthesise antifreeze proteins which can effectively inhibit the growth of ice crystals. The mechanism relies on adsorption of these proteins to the ice surface. Ellipsometry has been used to quantify glycopeptide antifreeze adsorption to the basal and prism faces of single ice crystals. The rate of accumulation was determined as a function of time and at concentrations between 0.0005 and 1.2 mg/ml. Estimates of packing density at saturation coverage have been made for the basal and prism faces.     

采用地统计学方法对水曲柳人工纯林表层根量的估计

采用地统计学的变异函数分析方法定量研究了水曲柳纯林表层根量的空间异质性特征,利用地统计学的克里格内插法结合定积分,对水曲柳纯林表层(0～10 cm)各类型根量进行了估测。结果表明:1)水曲柳细根(<2 mm)、粗根(≥2 mm)、活根和土壤总根变异函数曲线的理论模型符合球状模型,其它植物活根符合指数模型,水曲柳及其它植物死根符合线性模型。水曲柳及其它植物死根的空间变异主要是由随机性因素引起;水曲柳细根、粗根、活根、其它植物活根和土壤总根的空间变异主要是由结构性因素引起,且空间自相关程度均属中等(空间结构比在2 5 %和75 %之间)。水曲柳粗根的空间变异尺度为13.2 m,水曲柳细根、水曲柳活根、土壤总根和其它植物活根的空间变异尺度均大于33.9m。根据理论模型的拟合参数估计,水曲柳细根、水曲柳活根和土壤总根的空间变异尺度为6 1m,其它植物活根为183m。(2 )平均值成对二样本t检验结果表明,变异函数分析结果基础上的克里格内插法适用于水曲柳样地各空间位置处各类型根量的估计。利用此估计值,拟合其与位置坐标值之间的多元回归关系均为二元十次余弦级数多项式。利用此多项式,通过定积分的方法(积分区间为整块样地的大小) ,估计出水曲柳纯林表层根量为1.5 92 6 t/ hm2 ,水曲柳细根为0 .6 86 5 t/     

Hepatic iodothyronine 5'-deiodinase. The role of selenium.

Selenium (Se) deficiency decreased by 8-fold the activity of type 1 iodothyronine 5'-deiodinase (ID-I) in hepatic microsomal fractions from rats. Solubilized hepatic microsomes from rats injected with 75Se-labelled Na2SeO3 4 days before killing were found by chromatography on agarose gels to contain a 75Se-containing fraction with ID-I activity. PAGE of this fraction under reducing conditions, followed by autoradiography, revealed a single 75Se-containing protein (Mr 27,400 +/- 300). This protein could also be labelled with 125I-bromoacetyl reverse tri-iodothyronine, an affinity label for ID-I. The results suggest that hepatic ID-I is a selenoprotein or has an Se-containing subunit essential for activity.     

The effect of complex-formation with polyanions on the redox properties of cytochrome c.

1. The stable complex formed between mammalian cytochrome c and phosvitin at low ionic strength was studied by partition in an aqueous two-phase system. Oxidized cytochrome c binds to phosvitin with a higher affinity than reduced cytochrome c. The difference was equivalent to a decrease of the redox potential by 22 mV on binding. 2. Complex-formation with phosvitin strongly inhibited the reaction of cytochrome c with reagents that react as negatively charged species, such as ascorbate, dithionite, ferricyanide and tetrachlorobenzoquinol. Reaction with uncharged reagents such as NNN'N'-tetramethylphenylenediamine and the reduced form of the N-methylphenazonium ion (present as the methylsulphate) was little affected by complex-formation, whereas oxidation of the reduced cytochrome by the positively charged tris-(phenanthroline)cobalt(III) ion was greatly stimulated. 3. A similar pattern of inhibition and stimulation of reaction rates was observed when phosvitin was replaced by other macromolecular polyanions such as dextran sulphate and heparin, indicating that the results were a general property of complex-formation with polyanions. A weaker but qualitatively similar effect was observed on addition of inositol hexaphosphate and ATP. 4. It is suggested that the effects of complex-formation with polyanions on the reactivity of cytochrome c with redox reagents are mainly the result of replacing the positive charge on the free cytochrome by a net negative charge. Any steric effects on polyanion binding are small in comparison with such electrostatic effects.     

Activation of ADP-ribosyltransferase in polyamine-depleted mammalian cells.

Mammalian fibroblasts were cultured in the presence of alpha-methylornithine and/or methylglyoxal bis(guanylhydrazone), which inhibit the synthesis of polyamines. This led to a decrease in the cellular content of the polyamines spermine and spermidine by up to 60% when the cells were grown in the presence of both drugs together. The activity of the chromatin-associated enzyme ADP-ribosyltransferase was enhanced 2-3-fold in the drug-treated cells when measured in cells subsequently rendered permeable to exogenous NAD+, the substrate for the transferase. This is a novel and surprising observation, since the transferase is invariably activated by the addition of polyamines to a suitable incubation system such as permeabilized cells, isolated nuclei or the purified enzyme. We found no evidence that the activation was due to the appearance of DNA strand breaks, by using a variety of procedures including both neutral [the 'nucleoid' technique of Cook & Brazell [(1975) J. Cell Sci. 19, 261-279; (1976) J. Cell Sci. 22, 287-302]] and alkaline sucrose-gradient centrifugation and gel electrophoresis, suggesting that this therefore may not be the only means of regulating the activity of ADP-ribosyltransferase and that polyamines may have a role to play in this regard in vivo.     

Cloning of a coconut endosperm cDNA encoding a 1-acyl-sn-glycerol-3-phosphate acyltransferase that accepts medium-chain-length substrates.

Immature coconut (Cocos nucifera) endosperm contains a 1-acyl-sn-glycerol-3-phosphate acyltransferase (LPAAT) activity that shows a preference for medium-chain-length fatty acyl-coenzyme A substrates (H.M. Davies, D.J. Hawkins, J.S. Nelsen [1995] Phytochemistry 39:989-996). Beginning with solubilized membrane preparations, we have used chromatographic separations to identify a polypeptide with an apparent molecular mass of 29 kD, whose presence in various column fractions correlates with the acyltransferase activity detected in those same fractions. Amino acid sequence data obtained from several peptides generated from this protein were used to isolate a full-length clone from a coconut endosperm cDNA library. Clone pCGN5503 contains a 1325-bp cDNA insert with an open reading frame encoding a 308-amino acid protein with a calculated molecular mass of 34.8 kD. Comparison of the deduced amino acid sequence of pCGN5503 to sequences in the data banks revealed significant homology to other putative LPAAT sequences. Expression of the coconut cDNA in Escherichia coli conferred upon those cells a novel LPAAT activity whose substrate activity profile matched that of the coconut enzyme.     

Engineering the substrate specificity of rhizopuspepsin: the role of Asp 77 of fungal aspartic proteinases in facilitating the cleavage of oligopeptide substrates with lysine in P1.

Rhizopuspepsin and other fungal aspartic proteinases are distinct from the mammalian enzymes in that they are able to cleave substrates with lysine in the P1 position. Sequence and structural comparisons suggest that two aspartic acid residues, Asp 30 and Asp 77 (pig pepsin numbering), may be responsible for generating this unique specificity. Asp 30 and Asp 77 were changed to the corresponding residues in porcine pepsin, Ile 30 and Thr 77, to create single and double mutants. The zymogen forms of the wild-type and mutant enzymes were overexpressed in Escherichia coli as inclusion bodies. Following solubilization, denaturation, refolding, activation, and purification to homogeneity, structural and kinetic comparisons were made. The mutant enzymes exhibited a high degree of structural similarity to the wild-type recombinant protein and a native isozyme. The catalytic activities of the recombinant proteins were analyzed with chromogenic substrates containing lysine in the P1, P2, or P3 positions. Mutation of Asp 77 resulted in a loss of 7 kcal mol-1 of transition-state stabilization energy in the hydrolysis of the substrate containing lysine in P1. An inhibitor containing the positively charged P1-lysine side chain inhibited only the enzymes containing Asp 77. Inhibition of the Asp 77 mutants of rhizopuspepsin and several mammalian enzymes was restored upon acetylation of the lysine side chain. These results suggest that an exploitation of the specific electrostatic interaction of Asp 77 in the active site of fungal enzymes may lead to the design of compounds that preferentially inhibit a variety of related Candida proteinases in immunocompromised patients.     

昆虫多样性参数的测定和表达

简要介绍了文献中常见的表示昆虫多样性的3个指数物种数、辛普森指数和香农-维纳指数的测定和计算方法,讨论了香农-维纳指数和均匀度的英文字符的正确表达形式。     

The P700-chlorophyll a-protein of a blue-green alga

The previously isolated chlorophyll a-protein of blue-green algae has been shown to contain P700 in a ratio of 1 reaction center molecule per 100 light-harvesting chlorophyll molecules. One-fifth of the molecules in the preparation contain P700 together with some 20 light-harvesting molecules, whereas the other molecules contain bulk chlorophyll only. Both pigment-protein entities are considered to be essentially the same and cannot be fractionated. An aggregate containing both types probably makes up the photochemical portion of the algal Photosystem I in vivo. The absorption and emission spectra of the pigment-protein are reported, as well as the spectral changes associated with the photochemical reaction. In addition to chlorophyll, carotenoid and protein the complex contains a quinone, which is not a plastoquinone. This unidentified quinone appears to participate in secondary electron transfer reactions occurring in the complex. Horse cytochrome c can be bound to the complex and will donate electrons to P⁺700 upon illumination. Current hypotheses for the identity of the primary electron acceptor were tested. It appears unlikely that flavins, pteridines or iron fill this role.