The voltage-gated potassium channel Kv1.3 is an attractive target for treatment of autoimmune diseases. ShK toxin from sea anemone is one of the most potent blockers of Kv1.3, and therefore ShK and its analogues have been proposed as therapeutic leads for such diseases. Increasing the selectivity of the proposed leads for Kv1.3 over other Kv1 channels is a major issue in this endeavor. Here we study binding of ShK toxin to Kv1 channels using free energy simulation methods. Homology models for Kv1.1 and Kv1.3 channels are constructed using the crystal structure of Kv1.2. The initial poses for the Kv1.x-ShK complexes are obtained using HADDOCK, which are then refined via molecular dynamics simulations. The binding mode in each complex is characterized by identifying the strongly interacting residues, which compare well with available mutagenesis studies. For each complex, the potential of mean force is calculated from umbrella sampling simulations, and the corresponding absolute binding free energy is determined. The computed binding free energies are in good agreement with the experimental data, which increases the confidence on the model complexes. The insights gained on Kv1.x-ShK binding modes will be valuable in the development of new ShK analogues with better selectivity properties.

Combined efforts of farmers, rice scientists, extension personnel and Government of Bangladesh have yielded clean rice growth rate of 0.34 million ton (MT) year -1 during 2009-10 to 2013-14 in the country. In 2014-15, the country acquired a rice surplus of about 2 MT. However, maintaining the current surplus of rice in the coming decades is a great challenge. Authentic estimation of future rice requirement and future resource availability would guide to way forward. This paper presents rice vision for Bangladesh leading to 2050 and beyond. In this study, secondary data from different government-owned statistics and research institutes were collected, analyzed and synthesized to develop models and/or model parameters to generate outputs such as future population, rice production and rice requirement. Population of Bangladesh will reach 215.4 million in 2050, when 44.6 MT of clean rice will be required. With the pace of rice-production-increase in the last five years, production can reach 47.2 MT, having a surplus of 2.6 MT in 2050. The study sets 2.6 MT as the target for clean rice surplus every year leading to 2050 and beyond. Several hurdles, such as increasing population, decreasing resources and increasing climate vulnerability, can hinder achieving the target. Three major interventions-accelerating genetic gain, minimizing yield gap and curtailing adoption lag-are proposed to break the barriers to achieve the target. Major challenges to implement the interventions include shrinking net cropped area, decreasing availability of irrigation water and increasing pressure on soil fertility. Smart technology such as, location specific variety, profitable cropping sequences, innovative cultural management, and mechanization coupled with smart dissemination using multiple means would ease production barriers. We recommend a number of measures, such as, guaranteeing a minimum cropped area, accelerating the rate of genetic gain in varietal development and intensifying collaboration among the stakeholders to reduce adoption lag of newly released promising rice varieties, to achieve the rice vision of Bangladesh leading to 2050 and beyond.

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The voltage-gated potassium channel Kv1.3 is a well-established target for treatment of autoimmune diseases. ShK peptide from a sea anemone is one of the most potent blockers of Kv1.3 but its application as a therapeutic agent for autoimmune diseases is limited by its lack of selectivity against other Kv channels, in particular Kv1.1. Accurate models of Kv1.x-ShK complexes suggest that specific charge mutations on ShK could considerably enhance its specificity for Kv1.3. Here we evaluate the K18A mutation on ShK, and calculate the change in binding free energy associated with this mutation using the path-independent free energy perturbation and thermodynamic integration methods, with a novel implementation that avoids convergence problems. To check the accuracy of the results, the binding free energy differences were also determined from path-dependent potential of mean force calculations. The two methods yield consistent results for the K18A mutation in ShK and predict a 2 kcal/mol gain in Kv1.3/Kv1.1 selectivity free energy relative to wild-type peptide. Functional assays confirm the predicted selectivity gain for ShK[K18A] and suggest that it will be a valuable lead in the development of therapeutics for autoimmune diseases.

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ShK, a 35-residue peptide from a sea anemone, is a potent blocker of potassium channels. Here we describe a new ShK analogue with an additional C-terminus Lys residue and amide. ShK-K-amide is a potent blocker of Kv1.3 and, in contrast to ShK and ShK-amide, is selective for Kv1.3. To understand this selectivity, we created complexes of ShK-K-amide with Kv1.3 and Kv1.1 using docking and molecular dynamics simulations, then performed umbrella sampling simulations to construct the potential of mean force of the ligand and calculate the corresponding binding free energy for the most stable configuration. The results agree well with experimental data.

The voltage-gated potassium channel Kv1.3 is an established target for treatment of autoimmune diseases. Hence, there are intense efforts to develop immunosuppressant drugs from Kv1.3 blockers. ShK toxin from sea anemone is the most advanced peptide in this regard, but its lack of selectivity for Kv1.3 over Kv1.1 is an ongoing concern. The scorpion toxin HsTx1 is an equally potent blocker of Kv1.3, which is also selective for Kv1.3. It is of interest to understand the molecular basis of this selectivity as the lessons learned may suggest new avenues for enhancing the selectivity of other Kv1.3 blockers. Here we construct accurate models of Kv1.x-HsTx1 complexes using docking and molecular dynamics simulations. For each complex, the binding free energy of HsTx1 is determined from the potential of mean force calculations. Good agreement is found between the computed and experimental binding free energies, which increases confidence in the complex models. Comparison of the binding modes of HsTx1 with Kv1.1 and Kv1.3 reveals that the lower affinity of HsTx1 for Kv1.1 is due to its inability to come close to the pore domain, which prevents the pore inserting lysine from making proper contacts with the tyrosine carbonyls in the selectivity filter.

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