Synthetic peptides including acidic clusters as substrates and inhibitors of rat liver casein kinase TS (type-2)

The hexapeptides AcSer-Glu-Glu-Glu-Val-Glu and Ser-Glu-Glu-Glu-Glu-Glu, reminiscent of the sites phosphorylated by type-2 casein kinase TS in troponin T and glycogen synthase, respectively, have been synthesized and tested as phosphorylatable substrates for casein kinase TS as well as for other protein kinases. Both peptides are readily phosphorylated by casein kinase TS but not, to any detectable extent, by either cAMP-dependent protein kinase or phosphorylase kinase. Phosphorylation by type-1 casein kinase S was almost negligible. On the other hand the hexapeptide Ser-Glu-Glu-Glu-Ala-Ala is phosphorylated much more slowly and the hexapeptide Ser-Glu-Glu-Ala-Ala-Ala is almost unaffected by casein kinase TS. While the Vmax values of casein kinase TS with the acidic hexapeptides are comparable to those obtained with the corresponding protein substrates, the apparent Km values for the peptides are about two orders of magnitude higher than those for the protein substrates. The heptapeptide Arg-Ser-Glu-Glu-Glu-Val-Glu is a very poor substrate of casein kinase TS in comparison with the corresponding hexapeptide lacking the N-terminal Arg; it is, however, a competitive inhibitor toward the protein substrates, exhibiting a Ki similar to those of Ser-Glu-Glu-Glu-Glu-Glu and (Glu)5 which, in turn, are one order of magnitude higher than that of (Glu)10. It is concluded that the minimum structural requirement of type-2 casein kinases consists of a phosphorylatable residue followed by an acidic cluster, whose length is critical for the binding to the enzyme. Additional residues on the N-terminal side are not required, but their nature can influence the transphosphorylation reaction considerably.     

Exposure of Dunnocks (Prunella modularis) to their Previous Wintering Site Modifies Autumnal Activity Pattern: Evidence for Site Recognition?

We caught dunnocks at a wintering site near Pisa/Italy prior to their departure for breeding territories and held them indoors north of this site (Andechs/Germany) on a simulated photoperiod of 52.5 °N. After birds had gone through a reproductive cycle and postnuptial moult they developed migratory restlessness in autumn. At this time one group was transferred back to the previous wintering site (Pisa) where birds were held in individual activity cages in an outdoor aviary, allowing them to perceive as much environmental information as possible. A second group was transferred to a control site near Tour du Valat/France of approximately the same latitude and climate, but different longitude and held in an identical aviary. The diurnal activity pattern changed after transfer back to the previous wintering site, but not after transfer to the control site. Specifically, the amount of morning activity was reduced while afternoon activity was increased. This effect was restricted to those individuals that had been developing nocturnal migratory restlessness the previous spring. It was absent in individuals without migratory restlessness in spring, indicating that the different patterns were not due to unspecific effects from the testing sites. These results are consistent with the hypothesis that birds were able to derive information about their locality and to recognize their previous wintering site, resulting in suppression of migratory state by experience. The results are not definitely conclusive, however, because of several difficulties in the interpretation of perch-hopping activity, which are discussed in detail.     

Conformation and ion binding properties of peptides related to calcium binding domain III of bovine brain calmodulin

The conformational and ion binding properties of the sequences 93-104, 96-104, and 93-98 of domain III of bovine brain calmodulin (CaM) have been studied by CD and Tb3+-mediated fluorescence. In aqueous solution the interaction of all fragments with Ca2+ and Mg2+ ions is very weak and without any effect on the peptide conformation, which remains always random. In trifluoroethanol the interaction is very strong and the different fragments exhibit very distinct binding properties. In particular, the dodecapeptide fragment 93-104, and its N-terminal hexapeptide 98-104, bind calcium and magnesium with a very high binding constant (Kb greater than 10(5) M-1), undergoing a substantial conformational change. The structural rearrangement is particularly evident in the hexapeptide fragment, which tend to form a beta-bend. The C-terminal nonapeptide fragment 96-104 interacts with calcium and magnesium more weakly, and the binding process causes a decrease of ordered structure. These results suggest that, even in the entire dodecapeptide sequence corresponding to the loop of domain III of CaM, the calcium binding site is shifted toward the N-terminal hexapeptide segment. This interpretation is consistent with the results of crystallographic studies of CaM, which show that the calcium ions are located toward the amino terminal portion of the loop.     

Scanning and transmission electron microscopy evidence of the cytological effect of pyrrolnitrin on ‘Microsporon audouinii’ Gruby CBS 313-54 ‘in vitro’

Scanning and transmission electron microscopy showed that a ceiling quantity (1.56 mcg) of antifungal antibiotic Pyrrolnitrin caused heavy damage to dermathophyteMicrosporon audouinii Gruby CBS 313-54in vitro. Suitable preparation technique made it clear that the changes involved consisted of hyphal collapse on the edge of the culture, with loss of euplasmic organelles identity and cell autolysis. The cell wall, however, was apparently undamaged. These findings fit in with the suggestion that the mode of action of the antibiotic leads to generalised lipoproteic membranes damage. They must, however, be considered as representing the result of the terminal phase of cell distress.     

Environmental factors affecting the lipid metabolism in Rosmarinus officinalis L

The effect of some environmental factors on the lipid metabolism was studied in two chemotypes of Rosmarinus officinalis L. Epicuticular hydrocarbons (EH), epicuticular fatty acids (EFA), whole leaf fatty acids (WLFA) and essential oils (EO) were extracted and analysed by GC-MS during winter 1991–1992 and related to temperature and moisture variations. Leaf fresh and dry wts were determined along with some morphophysiological parameters such as specific leaf weight (SLW) and specific leaf area (SLA). Leaf areas were calculated by image analysis and statistically processed as for chemical data. The results indicated that in R. officinalis the response to some environmental factors, with particular reference to temperature and moisture, was an increase in epicuticular hydrocarbons and a decrease in epicuticular fatty acids, leaf fatty acids and essential oils. Qualitative changes in the chemical composition of the above lipid classes were found to be correlated with temperature changes. From a chemosystematic viewpoint, a clear separation between the two chemotypes was achieved only when epicuticular hydrocarbons and essential oils were considered as chemosystematic characters.     

Fishers' Knowledge Reveals Ecological Interactions Between Fish and Plants in High Diverse Tropical Rivers

Ecosystems - Frugivory and seed dispersal by fish is an important mutualistic interaction in complex and species-rich tropical rivers. The local ecological knowledge (LEK) held by fishers can...     

Physicochemical studies of amylose and its derivatives in aqueous solutions: Thermodynamics of the iodine–triiodide complex

Thermodynamic properties of the amylose–iodine–triiodide complex have been studied by spectrophotometry and by calorimetry using previously studied samples of amylose ionic derivatives, carboxymethylamylose and diethylaminoethylamylose. The ratio of triiodide to total molecular iodine ([I₃]_b/[I]_b + [I₂]_b) in the complex is ca. 0.3 over a range of iodide concentration from 10^?5 to 10^?4M, and there is no evidence for an increasing charge at slightly higher iodide concentration. Direct calorimetric experiments have been carried out in different conditions of polymer, iodine, and iodide concentration in order to study the dependence of the heat of the complexation as a function of the above parameters. It is shown that the dependence of the measured ΔH on the iodide concentration simply derives from the rearrangement of the triiodide equilibrium because of the uptake of a fixed ratio of iodine and triiodide molecules in the complex.     

Homing behaviour of pigeons disturbed by application of an olfactory stimulus

Summary Experiments designed to test the olfactory hypothesis of pigeon navigation by application of odorous substances to the birds' beaks and nostrils had shown until now variable results which have been interpreted differently. Using a new procedure, we were able to obtain consistent results. In each of the ten experiments performed, pigeons treated with -pinene were randomly oriented whereas control birds were not. Increase of homing time in experimental birds was also confirmed.This work was supported by a grant from the Consiglio Nazionale delle Ricerche.     

Conformational properties of the N-terminal residues of S-peptide. I. The ribonuclease S′ system

The contribution of the 1–6 N-terminal sequence to the conformational properties of the S-peptide (the 1–20 sequence of ribonuclease A) was assessed by determining in the ribonuclease S′ system the helical content and the binding capability of synthetic [Orn¹⁰]-S-peptide analogs, in which lysine¹, glutamic² and threonine³ were progressively deleted, alanine⁴ and alanine⁵were alternatively replaced by serine, and alanine⁶ was substituted by serine or proline. Both the deletion of the three N-terminal residues and the alanine⁶/proline replacement produces the loss of the helical structure up to lysine⁷. No or minor effects are found in all other cases. From the comparison of the binding data, the energy for the conformational stabilization of the N-terminal region was calculated to amount to 1.4 kcal/mol. The results are discussed in comparison with the known x-ray data of the enzyme, with some predictive rules of secondary structure which were applied to this region and with the known phylogenetic variance of the residues in this region.