Identification of a stability determinant on the edge of the Tet repressor four-helix bundle dimerization motif

Isofunctional tetracycline repressor (TetR) proteins isolated from different bacteria show a sequence identity between 38 and 88% of the residues. Their active state is a homodimer formed by a four-alpha-helix bundle as the main interaction motif. We utilize this sequence variation of isofunctional proteins to determine residues contributing to the stability of the four-helix bundle. The thermodynamic stabilities of two TetR proteins with 63% sequence identity were determined by urea-induced reversible denaturation followed by fluorescence and circular dichroism. Both methods yield identical results. The deltaG(o)U (H2O) values are 60 and 75 kJ x mol(-1). We have constructed TetR hybrid proteins derived from these wild types to identify the determinant leading to the 15 kJ x mol(-1) stability difference. Successive size reduction of the exchanged portion yielded two single residues affecting the overall protein stability. The P184Q exchange leads to a more stable protein, whereas the G181D exchange located at the solvent's exposed edge of the four-helix bundle is solely responsible for the reduced stability. Additional mutants based on crystal structures of TetR do not reveal any hint for steric interference of the Asp181 side chain with neighboring residues. Thus, this is an example for the role played by surface-exposed turn residues for the stability of four-helix bundles. We assume that the larger conformational flexibility of Gly and the reduction of the negative surface charge could favor formation of the turn on the edge of the four-helix bundle.     

Referate

Ohne Zusammenfassung     

The Effect of a Canine-assisted Activity on College Student Perceptions of Family Supports and Current Stressors

Using data from a previously published study on effects of a canine-assisted activity (CAA) on college student stress the week before final examinations, we examined whether participation in this activity had effects on perceptions of 1) family supports (i.e., emotional distance to family members and pets) and 2) current stressors. A total of 74 students completed the Family Life Space Diagram (FLSD), which uses an individual's structured drawings of distances between symbols of self and living entities, organizations, and stressors to reflect “emotional distances.” Participants were randomly assigned to order of CAA or FLSD, which was the intervention study control condition. Groups completed the FLSD after participating in CAA (Group A, n = 34) or prior to CAA (Group B, n = 40). Participants were primarily white (56.8%) females (75.7%) with a mean age of 19.38 years (SD = 1.75). Significant differences with large effect sizes were found for both groups in distances between 1) self-closest and self-average family member (Group A: t = 7.02, df = 33, p < 0.001, d = 1.205; Group B: t = 6.25, df = 39, p < 0.001, d = 0.987) and 2) self-closest personal stressor (t = 2.93, df = 18, p = 0.009, d = 1.311) and self-average personal stressor (t = 2.54, df = 18. p = 0.020, d = 1.138). In both cases, Group A (FLSD following CAA) placed personal stressors in closer proximity to self. Although CAA did not affect students’ current perceptions of family and pet relationships, the intervention may have increased their abilities to cope with personal stressors. Modified stress theory supports the proposition that positive emotions associated with CAA engage positive coping strategies, resulting in more positive perceptions of stressors.     

C-banding pattern and polymorphism of Aegilops caudata and chromosomal constitutions of the amphiploid T. aestivum — Ae. caudata and six derived chromosome addition lines

Summary C-banding patterns were analysed in 19 different accessions of Aegilops caudata (= Ae. markgrafii, = Triticum dichasians) (2n = 14, genomically CC) from Turkey, Greece and the USSR, and a generalized C-banded karyotype was established. Chromosome specific C-bands are present in all C-genome chromosomes, allowing the identification of each of the seven chromosome pairs. While only minor variations in the C-banding pattern was observed within the accessions, a large amount of polymorphic variation was found between different accessions. C-banding analysis was carried out to identify Ae. caudata chromosomes in the amphiploid Triticum aestivum cv Alcedo — Ae. caudata and in six derived chromosome addition lines. The results show that the amphiploid carries the complete Ae. Caudate chromosome complement and that the addition lines I, II, III, IV, V and VIII carry the Ae. caudata chromosome pairs B, C, D, F, E and G, respectively. One of the two SAT chromosome pairs (A) is missing from the set. C-banding patterns of the added Ae. caudata chromosomes are identical to those present in the ancestor species, indicating that these chromosomes are not structurally rearranged. The results are discussed with respect to the homoeologous relationships of the Ae. caudata chromosomes.     

Estrogen biosynthesis and its inhibition--a review

A short review is given about extragonadal sources of estrogens via androgen aromatization; the possible importance of this aromatization; the mechanism of aromatization, the inhibition of androgen aromatization by steroidal compounds; the importance and possible usefulness of such an inhibition.     

Preparation and Antiviral Activity of Several Deoxygenated Ribavirin and Tiazofurin Derivatives

Abstract

Ribavirin and tiazofurin, two nucleosides of known antiviral activity, have been transformed by previously reported methods to yield several deoxy,epoxy, or dideoxy analogues. The deoxygenated derivatives were evaluated for antiviral activity against a host of DNA and RNA viruses; however, no significant in vitro activity was detected.     

Discovery of the Inhibitory Effect of a Phosphatidylinositol Derivative on P-Glycoprotein by Virtual Screening Followed by In Vitro Cellular Studies

P-glycoprotein is capable of effluxing a broad range of cytosolic and membrane penetrating xenobiotic substrates, thus leading to multi-drug resistance and posing a threat for the therapeutic treatment of several diseases, including cancer and central nervous disorders. Herein, a virtual screening campaign followed by experimental validation in Caco-2, MDKCII, and MDKCII mdr1 transfected cell lines has been conducted for the identification of novel phospholipids with P-gp transportation inhibitory activity. Phosphatidylinositol-(1,2-dioctanoyl)-sodium salt (8∶0 PI) was found to significantly inhibit transmembrane P-gp transportation in vitro in a reproducible-, cell line-, and substrate-independent manner. Further tests are needed to determine whether this and other phosphatidylinositols could be co-administered with oral drugs to successfully increase their bioavailability. Moreover, as phosphatidylinositols and phosphoinositides are present in the human diet and are known to play an important role in signal transduction and cell motility, our finding could be of substantial interest for nutrition science as well.