Fabrication And Characterization Of A RFN/CDH Nano-scaled Ceramic Surface Via LbL Self-assembly And Prediction Subcellular Localization Of RFN/CDH

Part Ⅰ Prediction of protein subcellular localization based on the general form ofChou's pseudo amino acid compositionObjective: With the rapid increase of protein sequences in the post-genomic age, it ischallenging to develop accurate and automated methods for reliably and quickly predictingtheir subcellular localizations. Till now, many efforts have been tried, but most of whichused only a single algorithm. Methods: In this paper, we proposed an ensemble classifier ofKNN (k-nearest neighbor) and SVM (support vector machine) algorithms to predict thesubcellular localization of proteins. This ensemble classifier was developed by fusing manybasic individual classifiers through a voting system. Results: The overall predictionaccuracies by the one-versus-one strategy are78.17%,89.94%and75.55%for threebenchmark datasets of eukaryotic proteins and95.22%,93.47%and80.72%for the otherthree benchmark datasets of proteins, respectively. Conclusion: The improved predictionaccuracies reveal that the general form of Chou's pseudo amino acid composition helps topredict subcellular locations of proteins (FN, CDH and rFN/CDH).Part Ⅱ Fabrication and characterization of a recombinant fibronectin/cadherinnano-scaled ceramic surface via layer-by-layer self-assemblyObjective: The purpose of this study is to investigate the feasibility of fabricating arecombinant fibronectin/cadherin protein (rFN/CDH) bio-inspired surface vialayer-by-layer (LbL) self-assembly technique. Methods: The rFN/CDH was prepared usingstandard gene cloning and protein techniques described in our previous work. TheChi-rFN/CDH multilayered structures were fabricated by LbL self-assembly technique viaelectrostatic interactions. The layer structure was characterized by X-ray photoelectron spectroscopy (XPS), Fourier transforms infrared spectroscopy (FTIR), water contact anglemeasurement, and Scanning Electron Microscope (SEM) analysis respectively. Results: A1954bp-long gene-coding fragment and80KD protein of rFN/CDH were successfullyprepared. After pre-assembly, the hydrophilicity of the surface showed a best condition withthe12-layers coating of BCP-PEI-(rFN/CDH-Chi)5-rFN/CDH. The contact angles were45°with pure BCP and30.1°with12-layers coating. The result of XPS analysis showed anitrogen peak (binding energy of400eV) and a sulfur peak (binding energy of164eV),which confirmed the success of self-assembly. Typical SEM photomicrographs revealed nosignificant differences in morphology between pure BCP and LbL-modified BCP at amagnification of10,000×. Conclusion: rFN/CDH was assembled to a scaffold surface inhigh efficiency via LbL self-assembly technique.