Endosymbiosestudien an schilülausen VI. Die nicht in symbiose lebende gattung apiomorpha und ihre ungewöhnliche embryonalentwicklung

Ohne ZusammenfassungAus der von der Deutschen Forschungsgemeinschaft eingerichteten, der Symbioseforschung dienenden Arbeitsstatte in Porto d'Ischia (Napoli).     

Protein kinase C inhibition by calmodulin and its fragments

Inhibition of protein kinase C (PKC) by calmodulin is investigated and we describe the localization of inhibitory sequences within the calmodulin molecule. We present evidence that calmodulin inhibits PKC through an inhibition of the activation of PKC associated with lipid membranes: Binding of PKC to lipid vesicles is not affected, but activation is abolished. The potent calmodulin antagonist R24571 (calmidazol) did not affect the inhibition of PKC by calmodulin at concentrations up to 10^–5 M. Two tryptic fragments of calmodulin were isolated which inhibited PKC. They were only slightly less potent than intact calmodulin with an IC₅₀ of 6 µ M compared to 1 µ M of intact calmodulin. They were identified as Ser³⁸-Arg⁷⁴ and His¹⁰⁷-Lys¹⁴⁸. Each of the inhibiting fragments contains an intact Ca²⁺-binding domain with complete helix-loop-helix structure (EF hand). Other calmodulin peptides showed only weak inhibitory activity. Both fragments did not stimulate cAMP phosphodiesterase even at concentrations 100-fold higher than the calmodulin concentration needed for maximal stimulation. None of the fragments acted as a calmodulin antagonist.     

Choline acetyltransferase-like immunoreactivity in the brain of Drosophila melanogaster

Summary Using a monoclonal antibody selective for the acetylcholine (ACh)-synthesizing enzyme choline acetyltransferase (ChAT) of Drosophila melanogaster we find ChAT-like immunoreactivity in specific synaptic regions throughout the brain of Drosophila melanogaster apart from the lobes and the peduncle of the mushroom body and most of the first visual neuropile (lamina). Several anatomically well-defined central brain structures exhibit particularly strong binding. Characteristic differential staining patterns are observed for each of the four neuromeres of the optic lobes. Cell bodies appear not to bind this antibody. The prominent features of the distribution of ChAT-like immunoreactivity are paralleled by the distribution of acetylcholine hydrolyzing enzymatic activity as revealed by histochemical staining for acetylcholine esterase (AChE). These results are discussed in comparison with published data on enzyme distribution, choline uptake and ACh receptor binding in the nervous system of Drosophila melanogaster.     

2,3-Butanediol production by Enterobacter aerogenes in continuous culture: role of oxygen supply

Summary The influence of oxygen on growth and production of 2,3-butanediol and acetoin by Enterobacter aerogenes was studied in continuous culture. At all dilution rates (D) studied cell mass increased steadily with increasing oxygen uptake rate (OUR), hence the micro-aerobic cultivation was energy-limited. The biomass yield on oxygen increased with D which suggests that cells need more energy for maintenance functions at lower D. At each D an optimal OUR giving highest volumetric productivity for the sum of butanediol and acetoin was found. The optimal OUR increased with D. The occurrence of optimal OURs results from the various effects of O₂ on growth and specific productivity. The latter was found to be a linear function of the specific OUR irrespective of D. At optimal OUR the cells proved to have equal specific OURs and equal specific productivities representing a fixed relationship between fermentative and respiratory metabolism. The product yield based on glucose and corrected for biomass formation was 80%. A product concentration as high as 43 g/l was obtained at D =0.1 h^–1 while the volumetric productivity was the highest at D =0.28 h^–1 (5.6 g/l and hour). The findings further indicate that growth and product generation are obviously non-associated phenomena. Hence, high productivities may be achievable by cell recycling and cell immobilisation systems. Offprint requests to: W.-D. Deckwer     

Two centuries of spatial and temporal dynamics of freshwater fish introductions

Aim

Investigating major freshwater fish flows (translocations) between biogeographic regions and their temporal dynamics and also quantifying spatial patterns and temporal changes in the array of introduced species, and the emergence and distance between major donor and recipient regions.

Location

Global.

Time Period

1800–2020.

Major Taxa Studied

Freshwater fishes.

Methods

We analysed a global dataset on freshwater fish introductions (4241 events of 688 species). Freshwater fish flows were investigated with flow diagrams and χ² tests, while PERMANOVA (permutational multivariate analysis of variance) was used to test the association between species and regions and temporal shifts. Cluster analysis revealed major recipient areas and composition of the introduced species. Finally, changes in distances between donor and recipient sites were tested with PERMANOVA.

Results

The number of introductions between biogeographic regions mirrored the European and North American dominance before World War II (WWII) and the trends in recreational fishing, biocontrol programmes and food production, especially in the Sino-Oriental region, which has a long tradition of aquaculture and fishkeeping. Over the years, the origins and composition of introduced species changed uniquely in each biogeographic region, although the most introduced species are common to every region. Salmonids and other cold-water species were frequently introduced before the 1950s, whereas tropical ornamental and aquaculture species currently prevail. Distances between donor and recipient sites did not vary over the time. After WWII, the Sino-Oriental region consolidated its dominance and the Ethiopian and Neotropical regions emerged as new global donor and recipient regions.

Main Conclusions

Global policy should focus on tropical ornamental and aquaculture species, which could benefit from global warming, especially in the Sino-Oriental region, because it currently dominates freshwater fish species flows, and the Ethiopian and Neotropical regions, because they recently emerged as important global donor and recipient regions of freshwater fish introductions.     

Characterization of a quaternary-structured folding intermediate of an antibody Fab-fragment.

Antibody folding is a complex process comprising folding and association reactions. Although it is usually difficult to characterize kinetic folding intermediates, in the case of the antibody Fab fragment, domain-domain interactions lead to a rate-limiting step of folding, thus accumulating folding intermediates at a late step of folding. Here, we analyzed a late folding intermediate of the Fab fragment of the monoclonal antibody MAK 33 from mouse (kappa/IgG1). As a strategy for accumulation of this intermediate we used partial denaturation of the native Fab by guanidinium chloride. This denaturation intermediate, which can be populated to about 90%, is indistinguishable from a late-folding intermediate with respect to denaturation and renaturation kinetics. The spectroscopic analysis reveals a native-like secondary structure of this intermediate with aromatic side chains only slightly more solvent exposed than in the native state. The respective partner domains are weekly associated. From these data we conclude that the intramolecular association of the two chains during folding, with all domains in a native-like structure, follows a two-step mechanism. In this mechanism, presumably hydrophobic interactions are followed by rearrangements leading to the exact complementarity of the contact sites of the respective domains.