No abstract

Colchicine is a toxic alkaloid prevalent in autumn crocus (Colchicum autumnale) that binds to tubulin and inhibits polymerization of microtubules. Using combinatorial and rational protein design, we have developed an artificial binding protein based on the human lipocalin 2 that binds colchicine with a dissociation constant of 120 pm, i.e. 10000-fold stronger than tubulin. Crystallographic analysis of the engineered lipocalin, dubbed Colchicalin, revealed major structural changes in the flexible loop region that forms the ligand pocket at the open end of the eight-stranded β-barrel, resulting in a lid-like structure over the deeply buried colchicine. A cis-peptide bond between residues Phe71 and Pro72 in loop #2 constitutes a peculiar feature and allows intimate contact with the tricyclic ligand. Using directed evolution, we achieved an extraordinary dissociation half-life of more than 9 h for the Colchicalin-colchicine complex. Together with the chemical robustness of colchicine and availability of activated derivatives, this also opens applications as a general-purpose affinity reagent, including facile quantification of colchicine in biological samples. Given that engineered lipocalins, also known as Anticalin® proteins, represent a class of clinically validated biopharmaceuticals, Colchicalin may offer a therapeutic antidote to scavenge colchicine and reverse its poisoning effect in situations of acute intoxication.

The “Lakhta Center” is the headquarters of public joint stock company “Gazprom”, a key cultural and environmental project of Saint-Petersburg and the flagship object of the architectural, engineering and construc­tion industries in Russia. The facility was completed for operating and became a precedent for the implementa­tion of such a unique in scale and complexity project in Russia. It was realized on schedule and within the budg­et. The architectural dominant of the Lakhta Center is a tower with a height of 462 meters, which has become the tallest building in Europe. The following management challenges and solutions are under consideration in the distinctive paper: transfer of the object; land restrictions and related projects; the need to develop a regulatory documents; lack of relevant experience and competencies in Russia; building a flexible contracting system, sepa­ration of design and construction functions; a flexible approach to the preparation of project documentation, the main state expertise (Glavgosekspertiza) and the beginning of the zero cycle; building stages; package contract­ing system; managing by the global team; optimization of the project solutions; risk management system; global crisis; sanctions; changing the role of the customer / contractor during the project.