Применение метода AlphaScreen и AlphaLisa в разработке лекарственных препаратов и фармакокинетических исследованиях

   Drug development requires high-tech, simple, and sensitive methods. AlphaLISA method was announced as a universal method that would fit the listed criteria. However, research of other works on the pharmacokinetics of drugs related to this method showed a small number of pharmacokinetic studies in clinical trials. In this review, we focused on not only the positive aspects of the Alpha method, but also its disadvantages.   Разработка лекарственных препаратов нуждается в высокотехнологичных, простых и чувствительных методах. Метод AlphaLISA является универсальным методом, который подходил бы под перечисленные критерии. Однако анализ работ по фармакокинетике лекарственных препаратов, имеющих отношение к данному методу, демонстрирует незначительное количество фармакокинетических исследований при клинических испытаниях. В данной статье мы раскрываем не только положительные стороны метода Alpha, но и его недостатки

Competing Activities of Heterotrimeric G Proteins in Drosophila Wing Maturation

Drosophila genome encodes six alpha-subunits of heterotrimeric G proteins. The Gαs alpha-subunit is involved in the post-eclosion wing maturation, which consists of the epithelial-mesenchymal transition and cell death, accompanied by unfolding of the pupal wing into the firm adult flight organ. Here we show that another alpha-subunit Gαo can specifically antagonize the Gαs activities by competing for the Gβ13F/Gγ1 subunits of the heterotrimeric Gs protein complex. Loss of Gβ13F, Gγ1, or Gαs, but not any other G protein subunit, results in prevention of post-eclosion cell death and failure of the wing expansion. However, cell death prevention alone is not sufficient to induce the expansion defect, suggesting that the failure of epithelial-mesenchymal transition is key to the folded wing phenotypes. Overactivation of Gαs with cholera toxin mimics expression of constitutively activated Gαs and promotes wing blistering due to precocious cell death. In contrast, co-overexpression of Gβ13F and Gγ1 does not produce wing blistering, revealing the passive role of the Gβγ in the Gαs-mediated activation of apoptosis, but hinting at the possible function of Gβγ in the epithelial-mesenchymal transition. Our results provide a comprehensive functional analysis of the heterotrimeric G protein proteome in the late stages of Drosophila wing development

Kermit interacts with gαo, vang, and motor proteins in Drosophila planar cell polarity.

In addition to the ubiquitous apical-basal polarity, epithelial cells are often polarized within the plane of the tissue - the phenomenon known as planar cell polarity (PCP). In Drosophila, manifestations of PCP are visible in the eye, wing, and cuticle. Several components of the PCP signaling have been characterized in flies and vertebrates, including the heterotrimeric Go protein. However, Go signaling partners in PCP remain largely unknown. Using a genetic screen we uncover Kermit, previously implicated in G protein and PCP signaling, as a novel binding partner of Go. Through pull-down and genetic interaction studies, we find that Kermit interacts with Go and another PCP component Vang, known to undergo intracellular relocalization during PCP establishment. We further demonstrate that the activity of Kermit in PCP differentially relies on the motor proteins: the microtubule-based dynein and kinesin motors and the actin-based myosin VI. Our results place Kermit as a potential transducer of Go, linking Vang with motor proteins for its delivery to dedicated cellular compartments during PCP establishment