Interaction of Biliverdin Chromophore with Near-Infrared Fluorescent Protein BphP1-FP Engineered from Bacterial Phytochrome

Near-infrared (NIR) fluorescent proteins (FPs) designed from PAS (Per-ARNT-Sim repeats) and GAF (cGMP phosphodiesterase/adenylate cyclase/FhlA transcriptional activator) domains of bacterial phytochromes covalently bind biliverdin (BV) chromophore via one or two Cys residues. We studied BV interaction with a series of NIR FP variants derived from the recently reported BphP1-FP protein. The latter was engineered from a bacterial phytochrome RpBphP1, and has two reactive Cys residues (Cys15 in the PAS domain and Cys256 in the GAF domain), whereas its mutants contain single Cys residues either in the PAS domain or in the GAF domain, or no Cys residues. We characterized BphP1-FP and its mutants biochemically and spectroscopically in the absence and in the presence of denaturant. We found that all BphP1-FP variants are monomers. We revealed that spectral properties of the BphP1-FP variants containing either Cys15 or Cys256, or both, are determined by the covalently bound BV chromophore only. Consequently, this suggests an involvement of the inter-monomeric allosteric effects in the BV interaction with monomers in dimeric NIR FPs, such as iRFPs. Likely, insertion of the Cys15 residue, in addition to the Cys256 residue, in dimeric NIR FPs influences BV binding by promoting the BV chromophore covalent cross-linking to both PAS and GAF domains.Peer reviewe

Allosteric effects of chromophore interaction with dimeric near-infrared fluorescent proteins engineered from bacterial phytochromes

Fluorescent proteins (FPs) engineered from bacterial phytochromes attract attention as probes for in vivo imaging due to their near-infrared (NIR) spectra and use of available in mammalian cells biliverdin (BV) as chromophore. We studied spectral properties of the iRFP670, iRFP682 and iRFP713 proteins and their mutants having Cys residues able to bind BV either in both PAS (Cys15) and GAF (Cys256) domains, in one of these domains, or without these Cys residues. We show that the absorption and fluorescence spectra and the chromophore binding depend on the location of the Cys residues. Compared with NIR FPs in which BV covalently binds to Cys15 or those that incorporate BV noncovalently, the proteins with BV covalently bound to Cys256 have blue-shifted spectra and higher quantum yield. In dimeric NIR FPs without Cys15, the covalent binding of BV to Cys256 in one monomer allosterically inhibits the covalent binding of BV to the other monomer, whereas the presence of Cys15 allosterically promotes BV binding to Cys256 in both monomers. The NIR FPs with both Cys residues have the narrowest blue-shifted spectra and the highest quantum yield. Our analysis resulted in the iRFP713/Val256Cys protein with the highest brightness in mammalian cells among available NIR FPs.Peer reviewe

Temporal Correlations of Local Network Losses

We introduce a continuum model describing data losses in a single node of a packet-switched network (like the Internet) which preserves the discrete nature of the data loss process. {\em By construction}, the model has critical behavior with a sharp transition from exponentially small to finite losses with increasing data arrival rate. We show that such a model exhibits strong fluctuations in the loss rate at the critical point and non-Markovian power-law correlations in time, in spite of the Markovian character of the data arrival process. The continuum model allows for rather general incoming data packet distributions and can be naturally generalized to consider the buffer server idleness statistics

Genesis of the Russia’s economy innovative development

The innovative development of economy in Russia is possible only if the way of development will be based on the objective basic components, i.e. the corresponding system of premises and factors determining the innovative process. Namely the latter ensure a transformation of the fundamental processes into the application ones, as well as the dynamic development of the economy under the new trends and patterns influenceyesBelgorod State Universit

Investigation of the Influence of Antioxidant Compositions on Development of Microbiological Spoilage in Storage of Fruits

The studies are devoted to the scientific grounding of expedience of after-harvest processing by antioxidant compositions for preventing the development of pathogenic microflora on fruit surfaces during a long storage. For the studies were used apple fruits of the varieties Aidared, Golden Dushesse, Renet Simirenka, pear fruits of the varieties Victoria, Crimea Raisin and Cure, plum fruits of the varieties Voloshka, Stanley and Italian Ugorka. Fruits were processed by immersion in the following antioxidant compositions: ACM is a mixture of dimethyl sulfoxide, ionol and polyethylene glycols; AARL – mixture of ascorbic acid, routin and lecithin; DL – mixture of dimethyl sulfoxide, ionol and lecithin. Fruits, processed by water, were used as a control. Exposition - 10 seconds. Storage was carried out at the temperature 0±1 ºС, relative air humidity 90–95 %. It was established that in the period of fruits laying for storage, the mean amount of epiphyte microflora was fixed on surfaces of plump and pear fruits of the mean ripening term. In the variety composition of epiphyte microflora prevailed spores of mesophyl aerobic and facultative-anaerobic microorganisms. Their mean number on apple fruits surface was 9,6·103 CCU/g, pear fruits – 10,6 103 CCU/g, plump fruits – 18·103CCCU/g. AOC processing of all types of fruits essentially decreased the speed of both MAFAnM and micromycetes growth. It was demonstrated that the used compositions in 2…3,5 times decreased the level of day losses from microbiological spoilage during the whole storage period. The most positive effect was received at using compositions, based on dystinol and lecithin. Multifactor analysis determined that the level of day losses from microbiological spoilage was mainly influenced by factors of raw material variety features (factor A) and antioxidant compositions processing (factor D). The shares of influence are 24 and 21 % respectively

Evaluation of the Innovation Potential of the Regions of the Central Federal District of the Russian Federation

The present article analyzes the current approaches to the evaluation of innovation potential of the individual territories of the Russian Federation. The technique of evaluation of the innovation potential on the example of the Central Federal District regions by means of the functional model of evaluation is given consideration to. The algorithm of the innovation potential evaluation proposed by the authors includes six groups of indexes integrating twenty-eight individual indicator

q-Functional Wick's theorems for particles with exotic statistics

In the paper we begin a description of functional methods of quantum field theory for systems of interacting q-particles. These particles obey exotic statistics and are the q-generalization of the colored particles which appear in many problems of condensed matter physics, magnetism and quantum optics. Motivated by the general ideas of standard field theory we prove the q-functional analogues of Hori's formulation of Wick's theorems for the different ordered q-particle creation and annihilation operators. The formulae have the same formal expressions as fermionic and bosonic ones but differ by a nature of fields. This allows us to derive the perturbation series for the theory and develop analogues of standard quantum field theory constructions in q-functional form.Comment: 15 pages, LaTeX, submitted to J.Phys.

Exhausting domains of the symmetrized bidisc

We show that the symmetrized bidisc may be exhausted by strongly linearly convex domains. It shows in particular the existence of a strongly linearly convex domain that cannot be exhausted by domains biholomorphic to convex ones.Comment: 6 page

Regional aspects of sectoral digitalization : problems and prospects

Purpose: At present, high and sometimes record indicators have been achieved for the individual branches of agricultural production. Further intensive development of agriculture is due to the search for new ways to increase labor productivity and lower production costs which is the main aim of this research. Design/Methodology/Approach: The methodological basis served as a systematic approach, which ensured comprehensiveness and purposefulness. Analytical, abstract-logical, economic-statistical, monographic and experimental research methods were also used in this work to examine problems and prospects of sectoral digitalization. Findings: The agro-industrial complex demonstrates the growth of production indicators and is on the rise. The target indicators, laid down in the Food Security Doctrine, have already been achieved by many indices. To a large extent, it is due to a significant increase in the agro-industrial complex. Practical implications: A proposed platform that would ensure a breakthrough in further technical modernization of production, can be created only if the technologies are improved based on their digitalization and orientation towards a specific consumer. Originality/Value: The pin-point introduction of digital technologies in agricultural production confirms the high efficiency of the full innovative technical modernization. The proposed digital transformation of management in the agro-industrial complex of Russia is the value of this research with its originality compared to other ideas.peer-reviewe

Random Walks in Local Dynamics of Network Losses

We suggest a model for data losses in a single node of a packet-switched network (like the Internet) which reduces to one-dimensional discrete random walks with unusual boundary conditions. The model shows critical behavior with an abrupt transition from exponentially small to finite losses as the data arrival rate increases. The critical point is characterized by strong fluctuations of the loss rate. Although we consider the packet arrival being a Markovian process, the loss rate exhibits non-Markovian power-law correlations in time at the critical point.Comment: 4 pages, 2 figure