The Drosophila hugin gene codes for myostimulatory and ecdysis-modifying neuropeptides

In a genomic screen we isolated the Drosophila gene hugin (hug, cytology 87C1-2) by cross-hybridisation to a human glial cell line-derived neurotrophic factor cDNA. Upon cDNA sequence analysis and in vitro expression assays, the hugin gene was found to encode a signal peptide containing proprotein that was further processed in Schneider-2 cells into peptides similar to known neuropeptides. Two of the peptides were similar to FXPRL-amides (pyrokinins) and to the ecdysis-triggering hormone, respectively. The former displayed myostimulatory activity in a bioassay on the cockroach hyperneural muscle preparation, as well as in the Drosophila heart muscle assay. Hugin is expressed during the later half of embryogenesis and during larval stages in a subgroup of neurosecretory cells of the suboesophageal ganglion. Ubiquitous ectopic hugin expression resulted in larval death predominantly at or shortly after ecdysis from second to third instar, suggesting that at least one of the posttranslational cleavage products affects molting of the larva by interfering with the regulation of ecdysis.     

三种胰岛素在反相色谱上的保留行为与热力学性质

以C8 柱为固定相 ,5 4%～ 5 9%甲醇为流动相 ,在一个广泛的温度 10～ 6 5℃范围内 ,研究了人、牛、猪 3种生物的胰岛素在高效反相色谱上的保留行为和热力学性质。研究结果表明 3种胰岛素在结构上的细微差别 ,能在反相色谱行为上明显地显示出来。同时还测定了 3种胰岛素与C8 柱结合时的焓ΔHo 和熵ΔSo 变化情况 ,这些热力学参数 ,对多肽和疏水界面相互作用机制的研究具有重要的参考价值。实验结果证明 ,通过色谱热力学参数的测定 ,将为蛋白质折叠机制以及生物大分子相互作用机制的研究 ,提供一种极其有效的方法     

Inhibition and inactivation of bovine mammary and liver UDP-galactose-4-epimerases.

Bovine liver and mammary UDP-galactose-4-epimerases were investigated with respect to various inhibitors and inactivators. Uridine nucleotides and NADH are potent inhibitors with Ki values in the low micromolar range. The NAD+/NADH ratio may be an important physiological control mechanism for it affects markedly the activity of the enzyme with 50% inhibition occurring at a ratio of 20:1. In the presence of uridine nucleotides binding of NADH to the epimerases is enhanced. Consequently, the effect of changes in the NAD+/NADH ratio in vivo would not be immediately apparent as uridine nucleotides would slow down the displacement of NADH by NAD+. Neither uridine nor galactose 1-phosphate inhibits the purified enzymes as previously reported with the impure liver enzyme. Uridine nucleotides provide almost total protection against the apparent first order inactivation of the epimerases by trypsin and allow determination of dissociation constants. NAD+ partially protects against trypsin inactivation. Inactivation with various sulfhydryl reagents is complex and the results indicate that at least three sulfhydryl groups may be modified before total inactivation occurs. Partial inactivation occurs upon modification of the epimerases with 2-hydroxy-5-nitrogenzyl bromide. Some protection against this modification is provided by the combination of NAD+ and UDP.     

Circular dichroism studies on α- and β-ketosides of 5-acetamido-3,5-dideoxy- -glycero- -galacto-nonulopyranosonic acid (N-acetylneuraminic acid) and of some of its derivatives

The circular dichroism spectra of a number of N-acetylneuraminic acid derivatives in aqueous solution were studied. For all compounds, the Cotton effects were found to be in the spectral range of the acetamido and carboxyl chromophores. The c.d. curves of the methyl, ethyl, and allyl α- -ketosides are characterized by a broad, positive band centered at λ ≈ 195 nm with a slight skew towards the higher wavelengths and weak bands between λ 225 and 255 nm, whereas the methyl β- -ketoside and the corresponding methyl ester show only an intense positive band with a broad shoulder in the same spectral range. 5-Acetamido-3,5-dideoxy- -glycero-β- -galacto-nonulopyranose, its methyl β- -ketoside, and 5-acetamido-3,5-dideoxy- -glycero- -galacto-nonulopyranosonamide containing only the acetamido chromophore showed one single positive Cotton effect centered at λ ≈ 192 nm. The c.d. spectrum of 5-acetamido-3,5-dideoxy- -glycero- -galacto-nonulopyranosonic acid confirms the β- configuration of the free acid in aqueous solution, whereas the shape of the c.d. curve of O-(N-acetyl-α- -neuraminopyranosyl)-(2→3)-O-β- -galactopyranosyl-(1→4)- -glucopyranose resembles that of the methyl, ethyl, and allyl α- -ketosides 2-4.     

Photoproduction of molecular hydrogen from waste water of a sugar refinery by photosynthetic bacteria

Summary From enrichment cultures inoculated with water and sediments of a waste-water pond of a sugar refinery several photosynthetic nonsulphur bacteria have been isolated and tested for the ability to produce molecular hydrogen in the light. Strains have been found that utilize the freshly used, untreated waste substrate with higher yields than the laboratory strain used so far. Under the test conditions one strain showed higher hydrogen production rates from waste water than from any synthetic substrate.     

The effect of abscisic acid and K+ on xylem exudation from excised roots of Lupinus luteus

External application of abscisic acid (ABA) induces a relatively high rate of xylem exudation in excised roots of Lupinus luteus L. cv. Weiko III. The response is relatively slow with a lag period of ca 1 h. It is also slowly, but reversibly, abolished by application of 3.6 or 36 μ M cycloheximide (CHX). Contrary to expectation, K⁺ is not a significant factor in maintaining flow rates in lupin roots as no response was measured after adding K⁺ to root systems, from which K⁺ had been withheld for periods ranging from 3 h to several days. In fact, excised roots obtained from seedlings raised in the absence of K⁺ failed to respond to added K⁺. Total depletion of K⁺ is difficult to achieve, because of initial seed reserves, and prolonged exudation in lupins can be maintained utilising only a small proportion of the K⁺ originally present in the root tissue. Nevertheless, the data cast doubt on the general applicability of the concept that volume flow is maintained by an osmotic gradient with K⁺ as the principal mineral ion.     

TransCent: Computational enzyme design by transferring active sites and considering constraints relevant for catalysis

Background  

Computational enzyme design is far from being applicable for the general case. Due to computational complexity and limited knowledge of the structure-function interplay, heuristic methods have to be used.