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Fujie, T.

Publications and source records attributed to Fujie, T..

3 recordsLinked to original sources

Protocol for Membrane Permeability Prediction of Cyclic Peptides using Descriptors Obtained from Extended Ensemble Molecular Dynamics Simulations and Chemical Structures

Improving membrane permeability is crucial in cyclic peptide drug discovery. Although the approach based on molecular dynamics (MD) simulation is widely used, it is computationally expensive. Alternatively, machine learning can predict membrane permeability at negligible cost, but it requires a larger dataset. There are only 7991 experimental values of membrane permeability available at the newly developed database. Another challenge in predicting membrane permeability using machine learning arises from the unique stable conformation of each cyclic peptide, which is strongly related to membrane permeability but difficult to predict from chemical structure. Therefore, we developed a machine learning protocol using 3D descriptors obtained from MD simulations in addition to 2D descriptors obtained from chemical structure of cyclic peptides, respectively, to generate a universal model with a realistic computational cost. We targeted 252 peptides across four datasets and, to calculate their 3D descriptors, predicted their conformation outside the membrane, at the water/membrane interface, and in the membrane by MD simulations based on the replica exchange with solute tempering/replica exchange umbrella sampling method using 16 replicas. For machine learning, six different algorithms were used, ranging from simple methods such as ridge regression to more sophisticated methods such as XGBoost. The best prediction performance was obtained using XGBoost, with a Pearsons correlation coefficient R = 0.77 and root mean square error (RMSE) = 0.62. The important descriptors included those that describe the hydrophilicity and hydrophobicity of the peptide, conformational differences between inside and outside the membrane, and the degree of freedom of the peptide. We confirm the models ability to predict the membrane permeability of peptides that differ in chemical structure from the training data by predicting the external data consisting of 24 peptides and obtained R = 0.76, RMSE = 1.14. Furthermore, we extracted one of the four datasets of the training data, re-trained the model, and performed the prediction of the permeability coefficients of the extracted dataset. The results showed the models generic nature with R = 0.61 and RMSE = 0.74, when using position-specific (PS) 3D, and 2D descriptors. In such situations descriptors based on conformations obtained from MD are essential for the prediction.

biophysics↗

qMaLioffG: A single green fluorescent protein FLIM indicator enabling quantitative imaging of endogenous ATP

The widespread use of fluorescence lifetime imaging microscopy (FLIM) for quantitative imaging is hindered by the limited availability of a FLIM-based genetically encoded indicator using a conventional 488 nm laser. Here, we present qMaLioffG, a single green fluorescent protein FLIM indicator showing a fluorescence lifetime change in ATP concentration within the physiological range. This allows quantitative imaging of endogenous ATP to investigate cellular energy status of different cell types.

biophysics↗

Efficient differentiation and polarization of primary cultured neurons on poly(lactic acid) scaffolds with microgrooved structures

Synthetic biodegradable polymers including poly(lactic acid) (PLA) are attractive cell culture substrates because their surfaces can be micropatterned to support cell adhesion. The cell adhesion properties of a scaffold mainly depend on its surface chemical and structural features; however, it remains unclear how these characteristics affect the growth and differentiation of cultured cells or their gene expression. In this study, we fabricated two differently structured PLA nanosheets: flat and microgrooved. We assessed the growth and differentiation of mouse primary cultured cortical neurons on these two types of nanosheets after pre-coating with poly-D-lysine and vitronectin. Interestingly, prominent neurite bundles were formed along the grooves on the microgrooved nanosheets, whereas thin and randomly extended neurites were only observed on the flat nanosheets. Comparative RNA sequencing analyses revealed that the expression of genes related to postsynaptic density, dendritic shafts, and asymmetric synapses was significantly and consistently up-regulated in cells cultured on the microgrooved nanosheets when compared with those cultured on the flat nanosheets. These results indicate that microgrooved PLA nanosheets can provide a powerful means of establishing a culture system for the efficient and reproducible differentiation of neurons, which will facilitate future investigations of the molecular mechanisms underlying the pathogenesis of neurological disorders.

molecular biology↗