Jenn-Kang Hwang

Prediction of protein subcellular localization

Because the proteins function is usually related to its subcellular localization, the ability to predict subcellular localization directly from protein sequences will be useful for inferring protein functions. Recent years have seen a surging interest in the development of novel computational tools to predict subcellular localization. At present, these approaches, based on a wide range of algorithms, have achieved varying degrees of success for specific organisms and for certain localization categories. A number of authors have noticed that sequence similarity is useful in predicting subcellular localization. For example, Nair and Rost (Protein Sci 2002;11:2836–2847) have carried out extensive analysis of the relation between sequence similarity and identity in subcellular localization, and have found a close relationship between them above a certain similarity threshold. However, many existing benchmark data sets used for the prediction accuracy assessment contain highly homologous sequences—some data sets comprising sequences up to 80–90% sequence identity. Using these benchmark test data will surely lead to overestimation of the performance of the methods considered. Here, we develop an approach based on a two‐level support vector machine (SVM) system: the first level comprises a number of SVM classifiers, each based on a specific type of feature vectors derived from sequences; the second level SVM classifier functions as the jury machine to generate the probability distribution of decisions for possible localizations. We compare our approach with a global sequence alignment approach and other existing approaches for two benchmark data sets—one comprising prokaryotic sequences and the other eukaryotic sequences. Furthermore, we carried out all‐against‐all sequence alignment for several data sets to investigate the relationship between sequence homology and subcellular localization. Our results, which are consistent with previous studies, indicate that the homology search approach performs well down to 30% sequence identity, although its performance deteriorates considerably for sequences sharing lower sequence identity. A data set of high homology levels will undoubtedly lead to biased assessment of the performances of the predictive approaches—especially those relying on homology search or sequence annotations. Our two‐level classification system based on SVM does not rely on homology search; therefore, its performance remains relatively unaffected by sequence homology. When compared with other approaches, our approach performed significantly better. Furthermore, we also develop a practical hybrid method, which combines the two‐level SVM classifier and the homology search method, as a general tool for the sequence annotation of subcellular localization. Proteins 2006.

Three-Dimensional Reconstruction of Brain-wide Wiring Networks in Drosophila at Single-Cell Resolution

BACKGROUND Animal behavior is governed by the activity of interconnected brain circuits. Comprehensive brain wiring maps are thus needed in order to formulate hypotheses about information flow and also to guide genetic manipulations aimed at understanding how genes and circuits orchestrate complex behaviors. RESULTS To assemble this map, we deconstructed the adult Drosophila brain into approximately 16,000 single neurons and reconstructed them into a common standardized framework to produce a virtual fly brain. We have constructed a mesoscopic map and found that it consists of 41 local processing units (LPUs), six hubs, and 58 tracts covering the whole Drosophila brain. Despite individual local variation, the architecture of the Drosophila brain shows invariance for both the aggregation of local neurons (LNs) within specific LPUs and for the connectivity of projection neurons (PNs) between the same set of LPUs. An open-access image database, named FlyCircuit, has been constructed for online data archiving, mining, analysis, and three-dimensional visualization of all single neurons, brain-wide LPUs, their wiring diagrams, and neural tracts. CONCLUSION We found that the Drosophila brain is assembled from families of multiple LPUs and their interconnections. This provides an essential first step in the analysis of information processing within and between neurons in a complete brain.

Predicting subcellular localization of proteins for Gram‐negative bacteria by support vector machines based on n‐peptide compositions

Gram‐negative bacteria have five major subcellular localization sites: the cytoplasm, the periplasm, the inner membrane, the outer membrane, and the extracellular space. The subcellular location of a protein can provide valuable information about its function. With the rapid increase of sequenced genomic data, the need for an automated and accurate tool to predict subcellular localization becomes increasingly important. We present an approach to predict subcellular localization for Gram‐negative bacteria. This method uses the support vector machines trained by multiple feature vectors based on n‐peptide compositions. For a standard data set comprising 1443 proteins, the overall prediction accuracy reaches 89%, which, to the best of our knowledge, is the highest prediction rate ever reported. Our prediction is 14% higher than that of the recently developed multimodular PSORT‐B. Because of its simplicity, this approach can be easily extended to other organisms and should be a useful tool for the high‐throughput and large‐scale analysis of proteomic and genomic data.

KinasePhos 2.0: a web server for identifying protein kinase-specific phosphorylation sites based on sequences and coupling patterns

Due to the importance of protein phosphorylation in cellular control, many researches are undertaken to predict the kinase-specific phosphorylation sites. Referred to our previous work, KinasePhos 1.0, incorporated profile hidden Markov model (HMM) with flanking residues of the kinase-specific phosphorylation sites. Herein, a new web server, KinasePhos 2.0, incorporates support vector machines (SVM) with the protein sequence profile and protein coupling pattern, which is a novel feature used for identifying phosphorylation sites. The coupling pattern [XdZ] denotes the amino acid coupling-pattern of amino acid types X and Z that are separated by d amino acids. The differences or quotients of coupling strength CXdZ between the positive set of phosphorylation sites and the background set of whole protein sequences from Swiss-Prot are computed to determine the number of coupling patterns for training SVM models. After the evaluation based on k-fold cross-validation and Jackknife cross-validation, the average predictive accuracy of phosphorylated serine, threonine, tyrosine and histidine are 90, 93, 88 and 93%, respectively. KinasePhos 2.0 performs better than other tools previously developed. The proposed web server is freely available at http://KinasePhos2.mbc.nctu.edu.tw/.

Simulation of the dynamics of electron transfer reactions in polar solvents: Semiclassical trajectories and dispersed polaron approaches

Simulation methods for exploring the microscopic aspects of electron transfer (ET) reactions are developed. For the high temperature limit, where anharmonicity effects are crucial, we develop a semiclassical trajectory (ST) method. This method treats the reaction by considering the time dependence of the electronic energy gap along the classical trajectories of the solvent molecules. The ST approach, which appears as an ad hoc approach, might be at present the most rigorous practical models for simulating ET reactions. That is, this method reproduces the results of the quantum mechanical harmonic test case in the high temperature statistical limit. More importantly, for anharmonic systems it provides a rate constant which depends on the proper activation free energy; this dependence cannot be evaluated by any of the current quantum mechanical methods. For the low temperature range we develop a ‘‘dispersed polaron’’ model that uses the Fourier transform of the microscopic energy gap to evaluate the harmoni...

(PS)2: protein structure prediction server.

Protein structure prediction provides valuable insights into function, and comparative modeling is one of the most reliable methods to predict 3D structures directly from amino acid sequences. However, critical problems arise during the selection of the correct templates and the alignment of query sequences therewith. We have developed an automatic protein structure prediction server, (PS)2, which uses an effective consensus strategy both in template selection, which combines PSI-BLAST and IMPALA, and target–template alignment integrating PSI-BLAST, IMPALA and T-Coffee. (PS)2 was evaluated for 47 comparative modeling targets in CASP6 (Critical Assessment of Techniques for Protein Structure Prediction). For the benchmark dataset, the predictive performance of (PS)2, based on the mean GTD_TS score, was superior to 10 other automatic servers. Our method is based solely on the consensus sequence and thus is considerably faster than other methods that rely on the additional structural consensus of templates. Our results show that (PS)2, coupled with suitable consensus strategies and a new similarity score, can significantly improve structure prediction. Our approach should be useful in structure prediction and modeling. The (PS)2 is available through the website at .

Predicting RNA-binding sites of proteins using support vector machines and evolutionary information

BackgroundRNA-protein interaction plays an essential role in several biological processes, such as protein synthesis, gene expression, posttranscriptional regulation and viral infectivity. Identification of RNA-binding sites in proteins provides valuable insights for biologists. However, experimental determination of RNA-protein interaction remains time-consuming and labor-intensive. Thus, computational approaches for prediction of RNA-binding sites in proteins have become highly desirable. Extensive studies of RNA-binding site prediction have led to the development of several methods. However, they could yield low sensitivities in trade-off for high specificities.ResultsWe propose a method, RNAProB, which incorporates a new smoothed position-specific scoring matrix (PSSM) encoding scheme with a support vector machine model to predict RNA-binding sites in proteins. Besides the incorporation of evolutionary information from standard PSSM profiles, the proposed smoothed PSSM encoding scheme also considers the correlation and dependency from the neighboring residues for each amino acid in a protein. Experimental results show that smoothed PSSM encoding significantly enhances the prediction performance, especially for sensitivity. Using five-fold cross-validation, our method performs better than the state-of-the-art systems by 4.90%~6.83%, 0.88%~5.33%, and 0.10~0.23 in terms of overall accuracy, specificity, and Matthews correlation coefficient, respectively. Most notably, compared to other approaches, RNAProB significantly improves sensitivity by 7.0%~26.9% over the benchmark data sets. To prevent data over fitting, a three-way data split procedure is incorporated to estimate the prediction performance. Moreover, physicochemical properties and amino acid preferences of RNA-binding proteins are examined and analyzed.ConclusionOur results demonstrate that smoothed PSSM encoding scheme significantly enhances the performance of RNA-binding site prediction in proteins. This also supports our assumption that smoothed PSSM encoding can better resolve the ambiguity of discriminating between interacting and non-interacting residues by modelling the dependency from surrounding residues. The proposed method can be used in other research areas, such as DNA-binding site prediction, protein-protein interaction, and prediction of posttranslational modification sites.

The dynamics of the primary event in rhodopsins revisited

Abstract The conclusions obtained from an early simulation study of the primary event in the vision process are reexamined by simulating the dynamics of the primary photoisomerization reaction in bacteriorhodopsin using a detailed molecular model for the protein-chromophore complex. The calculated photoisomerization time is of the same order of magnitude as that predicted by the early simulation and found in recent experimental studies. Several simulations with different conditions indicate that the isomerization process involves an efficient transfer of energy from the reaction coordinate to other degrees of freedom and is better described by a damped-motion model than by an inertial model. The damped motion is expected to give a significant quantum yield for both the cis→trans and trans→cis cases only if the minimum of the excited state potential surface is located above the ground state maximum (unless the excited state surface is extremely shallow or if the inertial effects associated with the surface crossing process are very large). The present study suggests that the observed quantum yield should not be analyzed by one-dimensional models but by multidimensional microscopic simulations that consider the surface crossing process and the subsequent ground state relaxation processes.

Prediction of the bonding states of cysteines Using the support vector machines based on multiple feature vectors and cysteine state sequences

The support vector machine (SVM) method is used to predict the bonding states of cysteines. Besides using local descriptors such as the local sequences, we include global information, such as amino acid compositions and the patterns of the states of cysteines (bonded or nonbonded), or cysteine state sequences, of the proteins. We found that SVM based on local sequences or global amino acid compositions yielded similar prediction accuracies for the data set comprising 4136 cysteine‐containing segments extracted from 969 nonhomologous proteins. However, the SVM method based on multiple feature vectors (combining local sequences and global amino acid compositions) significantly improves the prediction accuracy, from 80% to 86%. If coupled with cysteine state sequences, SVM based on multiple feature vectors yields 90% in overall prediction accuracy and a 0.77 Matthews correlation coefficient, around 10% and 22% higher than the corresponding values obtained by SVM based on local sequence information. Proteins 2004;55:000–000.

CELLO2GO: a web server for protein subCELlular LOcalization prediction with functional gene ontology annotation.

CELLO2GO (http://cello.life.nctu.edu.tw/cello2go/) is a publicly available, web-based system for screening various properties of a targeted protein and its subcellular localization. Herein, we describe how this platform is used to obtain a brief or detailed gene ontology (GO)-type categories, including subcellular localization(s), for the queried proteins by combining the CELLO localization-predicting and BLAST homology-searching approaches. Given a query protein sequence, CELLO2GO uses BLAST to search for homologous sequences that are GO annotated in an in-house database derived from the UniProt KnowledgeBase database. At the same time, CELLO attempts predict at least one subcellular localization on the basis of the species in which the protein is found. When homologs for the query sequence have been identified, the number of terms found for each of their GO categories, i.e., cellular compartment, molecular function, and biological process, are summed and presented as pie charts representing possible functional annotations for the queried protein. Although the experimental subcellular localization of a protein may not be known, and thus not annotated, CELLO can confidentially suggest a subcellular localization. CELLO2GO should be a useful tool for research involving complex subcellular systems because it combines CELLO and BLAST into one platform and its output is easily manipulated such that the user-specific questions may be readily addressed.