Description de Falklandia gen.n. de I'Ocean Austral et définition des Lysianassoidea uristidiens (Crustacea, Amphipoda)

The revision of the antarctic–subantarctic species Orchomenopsis reducta Schellenberg, 1931, has led to its attribution to a new, highly apomorphic genus: Falklandia gen.n. A new definition of the uristid group is given and Falklandia with 36 other lysianassoid genera are attributed to this supposedly monophyletic group.     

Mapping and analysis of protein sulfhydryl groups by modification with 2-bromoacetamido-4-nitrophenol

A method for identifying cysteine-containing peptides in proteins is presented using 2-bromoacetamido-4-nitrophenol (BNP) to introduce an easily detectable probe. The formation of a covalent bond between the protein sulfhydryl group and the acetamido moiety of BNP introduces a chromophore with an absorbance maximum at 410 nm. The modified protein can then be cleaved with appropriate proteases and the resulting peptides separated by chromatographic methods. Monitoring the effluent at a single wavelength (405 nm) provides a rapid and simple method of detecting and isolating only those peptides which contain cysteine residue(s). The nitrophenol derivative is stable under conditions required for protease cleavage. The reagent is therefore useful for locating cysteine-containing peptides in protein digests and can be used to explore the accessibility of different cysteines under a variety of conditions. The ease of modification, specificity of reaction, product stability, and simple detection of modified peptides make BNP ideal for investigation of cysteine residues.     

Cat satellite DNA. Isolation using netropsin with CsCl gradients

The absence of centromeric bands in the karyotype of Felis catus is confirmed. It is also confirmed that no satellite band is visible in CsCl density gradients. However, a satellite is observed both by recentrifuging the fraction of the DNA that bands at high density in CsCl and by using netropsin to enhance the resolution of a CsCl gradient containing total F. catus DNA. The satellite, about 0.5% of total DNA, was isolated by repeated centrifugation in CsCl alone and in CsCl with netropsin. Netropsin was removed and a pure satellite DNA obtained. The reassociation kinetics (C0t1/2 less than 10(-3) M . s) show that the satellite is of the simple sequence type and hence a candidate for centromeric heterochromatin. Its cytological localisation awaits in situ hybridisation experiments.     

Comparison of cobalamin-independent and cobalamin-dependent methionine synthases from Escherichia coli: two solutions to the same chemical problem.

In Escherichia coli, two enzymes catalyze the synthesis of methionine from homocysteine using methyltetrahydrofolate as the donor of the required methyl group: cobalamin-dependent and cobalamin-independent methionine synthases. Comparison of the mechanisms of these two enzymes offers the opportunity to examine two different solutions to the same chemical problem. We initiated the research described here to determine whether the two enzymes were evolutionarily related by comparing the deduced amino acid sequences of the two proteins. We have determined the nucleotide sequence for the metE gene, encoding the cobalamin-independent methionine synthase. Our results reveal an absence of similarity between the deduced amino acid sequences of the cobalamin-dependent and cobalamin-independent proteins and suggest that the two have arisen by convergent evolution. We have developed a rapid one-step purification of the recombinant cobalamin-independent methionine synthase (MetE) that yields homogeneous protein in high yield for mechanistic and structural studies. In the course of these studies, we identified a highly reactive thiol in MetE that is alkylated by chloromethyl ketones and by iodoacetamide. We demonstrated that alkylation of this residue, shown to be cysteine 726, results in complete loss of activity. While we are unable to deduce the role of cysteine 726 in catalysis at this time, the identification of this reactive residue suggests the possibility that this thiol functions as an intermediate methyl acceptor in catalysis, analogous to the role of cobalamin in the reaction catalyzed by the cobalamin-dependent enzyme.     

Cyanide-Resistant Root Respiration and Tap Root Formation in Two Subspecies of Hypochaeris radicata

Root respiration of the tap root forming species Hypochaeris radicata L. was measured during tap root formation. A comparison was made of two subspecies: H. radicata L. ssp. radicata L., a subspecies from relatively rich soils, and H. radicata L. ssp. ericetorum Van Soest, a subspecies from poor acidic soils. Root respiration was high and to a large extent inhibited by hydroxamic acid (SHAM) before the start of the tap root formation, indicating a high activity of an alternative non-phosphorylative electron transport chain. The rate of root respiration was much lower and less sensitive to SHAM when a considerable tap root was present. However, root respiration was also cyanide-resistant when a tap root was present, indicating that the alternative pathway was still present. A decreased rate of root respiration coincided with an increase of the content of storage carbohydrates, mainly in the tap root. The level of reducing sugars was constant throughout the experimental period, and it was concluded that the activity of the alternative oxidative pathway was significant in oxidation of sugars that could not be utilized for purposes like energy production, the formation of intermediates for growth or for storage. Root respiration decreased after the formation of a tap root. This decrease could neither be attributed to a gradual disappearance of the alternative chain, nor to a decreased level of reducing sugars. No differences in respiratory metabolism between the two subspecies have been observed, suggesting that a high activity of the alternative oxidative pathway is not significant in adaptation of the present two subspecies to relatively nutrient-rich or poor soils.