Advanced Protein Modeling Method: Benchmarking and Applications in Computer-Aided Drug Discovery

Development of homology modeling methods will remain an area of active research. These methods aim to develop and model increasingly accurate three-dimensional structures of yet uncrystallized therapeutically relevant proteins e.g. Class A G-Protein Coupled Receptors. Incorporating protein flexibility is one way to achieve this goal. Here, I will discuss the enhancement and validation of the ligand-steered modeling, originally developed by Dr. Claudio Cavasotto, via cross modeling of the newly crystallized GPCR structures. This method uses known ligands and known experimental information to optimize relevant protein binding sites by incorporating protein flexibility. The ligand-steered models were able to model, reasonably reproduce binding sites and the co-crystallized native ligand poses of the β2 adrenergic and Adenosine 2A receptors using a single template structure. They also performed better than the choice of template, and crude models in a small scale high-throughput docking experiments and compound selectivity studies. Next, the application of this method to develop high-quality homology models of Cannabinoid Receptor 2, an emerging non-psychotic pain management target, is discussed. These models were validated by their ability to rationalize structure activity relationship data of two, inverse agonist and agonist, series of compounds. The method was also applied to improve the virtual screening performance of the β2 adrenergic crystal structure by optimizing the binding site using β2 specific compounds. These results show the feasibility of optimizing only the pharmacologically relevant protein binding sites and applicability to structure-based drug design projects.

Primary production at time series station DYFAMED

Phytoplankton carbon assimilation has been measured near monthly using the 14C method at DYFAMED France JGOFS time-series station from 1993 to 1999. Data were obtained using the "LET GO" technique, which allowed in situ injection of bicarbonate and incubation in enclosures at 10 depths. Incubation duration was 4 h around noon, from which daily production was estimated. The seasonal variation of the depth-integrated carbon assimilation exhibits a marked cycle. Maximum values reach 1.8 g C/m**2/d in March or April; constant lower values were observed from August to January, in the range 100-300 mg C/m**2/d. The annual primary production vary in the range 86-232 g C/m**2/yr, in the upper range of older estimations. Primary production normalized to chlorophyll a shows maximum values in the period of oligotrophy. This increase of carbon assimilation rate per unit of chlorophyll a appears as linked to the period of phosphorus-limited ecosystem, and vertical distribution of taxonomic pigments suggests a possible role of cyanobacteria. Potential export production has been estimated from primary production data and Fp ratio based on pigments concentrations. These estimates (which imply biological steady state conditions) vary in a wide range, from 19 to 71 g C/m**2/yr. There is a decoupling between years with high potential export production and years with high measured particulate fluxes, which highlights the question of balance by resupply of the limiting nutrients and the role of dissolved organic carbon. A possible shift of primary production towards a more regeneration-dominated system is suggested for recent years.

Abundance and biomass of dinoflagellates and ciliates at time series station Helgoland Roads, North Sea, 2007-2009

A monitoring programme for microzooplankton was started at the long-term sampling station ''Kabeltonne'' at Helgoland Roads (54°11.30' N; 7°54.00' E) in January 2007 in order to provide more detailed knowledge on microzooplankton occurrence, composition and seasonality patterns at this site and to complement the existing plankton data series. Ciliate and dinoflagellate cell concentration and carbon biomass were recorded on a weekly basis. Heterotrophic dinoflagellates were considerably more important in terms of biomass than ciliates, especially during the summer months. However, in early spring, ciliates were the major group of microzooplankton grazers as they responded more quickly to phytoplankton food availability. Mixotrophic dinoflagellates played a secondary role in terms of biomass when compared to heterotrophic species; nevertheless, they made up an intense late summer bloom in 2007. The photosynthetic ciliate Myrionecta rubra bloomed at the end of the sampling period. Due to its high biomass when compared to crustacean plankton especially during the spring bloom, microzooplankton should be regarded as the more important phytoplankton grazer group at Helgoland Roads. Based on these results, analyses of biotic and abiotic factors driving microzooplankton composition and abundance are necessary for a full understanding of this important component of the plankton.