Publications (14)53.09 Total impact
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Article: MOPED: Model Organism Protein Expression Database.
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ABSTRACT: Large numbers of mass spectrometry proteomics studies are being conducted to understand all types of biological processes. The size and complexity of proteomics data hinders efforts to easily share, integrate, query and compare the studies. The Model Organism Protein Expression Database (MOPED, htttp://moped.proteinspire.org) is a new and expanding proteomics resource that enables rapid browsing of protein expression information from publicly available studies on humans and model organisms. MOPED is designed to simplify the comparison and sharing of proteomics data for the greater research community. MOPED uniquely provides protein level expression data, meta-analysis capabilities and quantitative data from standardized analysis. Data can be queried for specific proteins, browsed based on organism, tissue, localization and condition and sorted by false discovery rate and expression. MOPED empowers users to visualize their own expression data and compare it with existing studies. Further, MOPED links to various protein and pathway databases, including GeneCards, Entrez, UniProt, KEGG and Reactome. The current version of MOPED contains over 43,000 proteins with at least one spectral match and more than 11 million high certainty spectra.Nucleic Acids Research 12/2011; 40(Database issue):D1093-9. · 8.03 Impact Factor -
Article: IPM: An integrated protein model for false discovery rate estimation and identification in high-throughput proteomics.
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ABSTRACT: In high-throughput mass spectrometry proteomics, peptides and proteins are not simply identified as present or not present in a sample, rather the identifications are associated with differing levels of confidence. The false discovery rate (FDR) has emerged as an accepted means for measuring the confidence associated with identifications. We have developed the Systematic Protein Investigative Research Environment (SPIRE) for the purpose of integrating the best available proteomics methods. Two successful approaches to estimating the FDR for MS protein identifications are the MAYU and our current SPIRE methods. We present here a method to combine these two approaches to estimating the FDR for MS protein identifications into an integrated protein model (IPM). We illustrate the high quality performance of this IPM approach through testing on two large publicly available proteomics datasets. MAYU and SPIRE show remarkable consistency in identifying proteins in these datasets. Still, IPM results in a more robust FDR estimation approach and additional identifications, particularly among low abundance proteins. IPM is now implemented as a part of the SPIRE system.Journal of proteomics 06/2011; 75(1):116-21. · 5.07 Impact Factor -
Article: SPIRE: Systematic protein investigative research environment.
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ABSTRACT: The SPIRE (Systematic Protein Investigative Research Environment) provides web-based experiment-specific mass spectrometry (MS) proteomics analysis (https://www.proteinspire.org). Its emphasis is on usability and integration of the best analytic tools. SPIRE provides an easy to use web-interface and generates results in both interactive and simple data formats. In contrast to run-based approaches, SPIRE conducts the analysis based on the experimental design. It employs novel methods to generate false discovery rates and local false discovery rates (FDR, LFDR) and integrates the best and complementary open-source search and data analysis methods. The SPIRE approach of integrating X!Tandem, OMSSA and SpectraST can produce an increase in protein IDs (52-88%) over current combinations of scoring and single search engines while also providing accurate multi-faceted error estimation. One of SPIRE's primary assets is combining the results with data on protein function, pathways and protein expression from model organisms. We demonstrate some of SPIRE's capabilities by analyzing mitochondrial proteins from the wild type and 3 mutants of C. elegans. SPIRE also connects results to publically available proteomics data through its Model Organism Protein Expression Database (MOPED). SPIRE can also provide analysis and annotation for user supplied protein ID and expression data.Journal of proteomics 05/2011; 75(1):122-6. · 5.07 Impact Factor -
Article: Technology and data-intensive science in the beginning of the 21st century.
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ABSTRACT: This article is a summary of the technology issues and challenges of data-intensive science and cloud computing as discussed in the Data-Intensive Science (DIS) workshop in Seattle, September 19-20, 2010.Omics: a journal of integrative biology 04/2011; 15(4):203-7. · 2.29 Impact Factor -
Article: The United States of America and scientific research.
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ABSTRACT: To gauge the current commitment to scientific research in the United States of America (US), we compared federal research funding (FRF) with the US gross domestic product (GDP) and industry research spending during the past six decades. In order to address the recent globalization of scientific research, we also focused on four key indicators of research activities: research and development (R&D) funding, total science and engineering doctoral degrees, patents, and scientific publications. We compared these indicators across three major population and economic regions: the US, the European Union (EU) and the People's Republic of China (China) over the past decade. We discovered a number of interesting trends with direct relevance for science policy. The level of US FRF has varied between 0.2% and 0.6% of the GDP during the last six decades. Since the 1960s, the US FRF contribution has fallen from twice that of industrial research funding to roughly equal. Also, in the last two decades, the portion of the US government R&D spending devoted to research has increased. Although well below the US and the EU in overall funding, the current growth rate for R&D funding in China greatly exceeds that of both. Finally, the EU currently produces more science and engineering doctoral graduates and scientific publications than the US in absolute terms, but not per capita. This study's aim is to facilitate a serious discussion of key questions by the research community and federal policy makers. In particular, our results raise two questions with respect to: a) the increasing globalization of science: "What role is the US playing now, and what role will it play in the future of international science?"; and b) the ability to produce beneficial innovations for society: "How will the US continue to foster its strengths?"PLoS ONE 01/2010; 5(8):e12203. · 4.09 Impact Factor -
Article: Development of BIATECH-54 standard mixtures for assessment of protein identification and relative expression.
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ABSTRACT: Mixtures of known proteins have been very useful in the assessment and validation of methods for high-throughput (HTP) MS (MS/MS) proteomics experiments. However, these test mixtures have generally consisted of few proteins at near equal concentration or of a single protein at varied concentrations. Such mixtures are too simple to effectively assess the validity of error rates for protein identification and differential expression in HTP MS/MS studies. This work aimed at overcoming these limitations and simulating studies of complex biological samples. We introduced a pair of 54-protein standard mixtures of variable concentrations with up to a 1000-fold dynamic range in concentration and up to ten-fold expression ratios with additional negative controls (infinite expression ratios). These test mixtures comprised 16 off-the-shelf Sigma-Aldrich proteins and 38 Shewanella oneidensis proteins produced in-house. The standard proteins were systematically distributed into three main concentration groups (high, medium, and low) and then the concentrations were varied differently for each mixture within the groups to generate different expression ratios. The mixtures were analyzed with both low mass accuracy LCQ and high mass accuracy FT-LTQ instruments. In addition, these 54 standard proteins closely follow the molecular weight distributions of both bacterial and human proteomes. As a result, these new standard mixtures allow for a much more realistic assessment of approaches for protein identification and label-free differential expression than previous mixtures. Finally, methodology and experimental design developed in this work can be readily applied in future to development of more complex standard mixtures for HTP proteomics studies.PROTEOMICS 11/2007; 7(20):3693-8. · 4.51 Impact Factor -
Article: Experiment-specific estimation of peptide identification probabilities using a randomized database.
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ABSTRACT: Determining the error rate for peptide and protein identification accurately and reliably is necessary to enable evaluation and crosscomparisons of high throughput proteomics experiments. Currently, peptide identification is based either on preset scoring thresholds or on probabilistic models trained on datasets that are often dissimilar to experimental results. The false discovery rates (FDR) and peptide identification probabilities for these preset thresholds or models often vary greatly across different experimental treatments, organisms, or instruments used in specific experiments. To overcome these difficulties, randomized databases have been used to estimate the FDR. However, the cumulative FDR may include low probability identifications when there are a large number of peptide identifications and exclude high probability identifications when there are few. To overcome this logical inconsistency, this study expands the use of randomized databases to generate experiment-specific estimates of peptide identification probabilities. These experiment-specific probabilities are generated by logistic and Loess regression models of the peptide scores obtained from original and reshuffled database matches. These experiment-specific probabilities are shown to very well approximate "true" probabilities based on known standard protein mixtures across different experiments. Probabilities generated by the earlier Peptide_Prophet and more recent LIPS models are shown to differ significantly from this study's experiment-specific probabilities, especially for unknown samples. The experiment-specific probabilities reliably estimate the accuracy of peptide identifications and overcome potential logical inconsistencies of the cumulative FDR. This estimation method is demonstrated using a Sequest database search, LIPS model, and a reshuffled database. However, this approach is generally applicable to any search algorithm, peptide scoring, and statistical model when using a randomized database.Omics A Journal of Integrative Biology 02/2007; 11(4):351-65. · 2.44 Impact Factor -
Article: Global profiling of Shewanella oneidensis MR-1: expression of hypothetical genes and improved functional annotations.
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ABSTRACT: The gamma-proteobacterium Shewanella oneidensis strain MR-1 is a metabolically versatile organism that can reduce a wide range of organic compounds, metal ions, and radionuclides. Similar to most other sequenced organisms, approximately 40% of the predicted ORFs in the S. oneidensis genome were annotated as uncharacterized "hypothetical" genes. We implemented an integrative approach by using experimental and computational analyses to provide more detailed insight into gene function. Global expression profiles were determined for cells after UV irradiation and under aerobic and suboxic growth conditions. Transcriptomic and proteomic analyses confidently identified 538 hypothetical genes as expressed in S. oneidensis cells both as mRNAs and proteins (33% of all predicted hypothetical proteins). Publicly available analysis tools and databases and the expression data were applied to improve the annotation of these genes. The annotation results were scored by using a seven-category schema that ranked both confidence and precision of the functional assignment. We were able to identify homologs for nearly all of these hypothetical proteins (97%), but could confidently assign exact biochemical functions for only 16 proteins (category 1; 3%). Altogether, computational and experimental evidence provided functional assignments or insights for 240 more genes (categories 2-5; 45%). These functional annotations advance our understanding of genes involved in vital cellular processes, including energy conversion, ion transport, secondary metabolism, and signal transduction. We propose that this integrative approach offers a valuable means to undertake the enormous challenge of characterizing the rapidly growing number of hypothetical proteins with each newly sequenced genome.Proceedings of the National Academy of Sciences 03/2005; 102(6):2099-104. · 9.68 Impact Factor -
Article: Charge state estimation for tandem mass spectrometry proteomics.
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ABSTRACT: High-throughput protein analysis by tandem mass spectrometry produces anywhere from thousands to millions of spectra that are being used for peptide and protein identifications. Though each spectrum corresponds only to one charged peptide (ion) state, repetitive database searches of multiple charge states are typically conducted since the resolution of many common mass spectrometers is not sufficient to determine the charge state. The resulting database searches are both error-prone and time-consuming. We describe a straightforward, accurate approach on charge state estimation (CHASTE). CHASTE relies on fragment ion peak distributions, and by using reliable logistic regression models, combines different measurements to improve its accuracy. CHASTE's performance has been validated on data sets, comprised of known peptide dissociation spectra, obtained by replicate analyses of our earlier developed protein standard mixture using ion trap mass spectrometers at different laboratories. CHASTE was able to reduce number of needed database searches by at least 60% and the number of redundant searches by at least 90% virtually without any informational loss. This greatly alleviates one of the major bottlenecks in high throughput peptide and protein identifications. Thresholds and parameter estimates can be tailored to specific analysis situations, pipelines, and instrumentations. CHASTE was implemented in Java GUI-based and command-line-based interfaces.Omics A Journal of Integrative Biology 02/2005; 9(3):233-50. · 2.44 Impact Factor -
Article: Global profiling of Shewanella oneidensis MR1: Expression of hypothetical genes and improved functional annotations
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ABSTRACT: The -proteobacterium Shewanella oneidensis strain MR-1 is a metabolically versatile organism that can reduce a wide range of organic compounds, metal ions, and radionuclides. Similar to most other sequenced organisms, 40% of the predicted ORFs in the S. oneidensis genome were annotated as uncharacterized "hypothetical" genes. We implemented an integrative approach by using experimental and computational analyses to provide more detailed insight into gene function. Global expression profiles were determined for cells after UV irradiation and under aerobic and suboxic growth conditions. Transcriptomic and proteomic analyses confidently identified 538 hypothetical genes as expressed in S. oneidensis cells both as mRNAs and proteins (33% of all predicted hypothetical proteins). Publicly available analysis tools and databases and the expression data were applied to improve the annotation of these genes. The annotation results were scored by using a seven-category schema that ranked both confidence and precision of the functional assignment. We were able to identify homologs for nearly all of these hypothetical proteins (97%), but could confidently assign exact biochemical functions for only 16 proteins (category 1; 3%). Altogether, computational and experimental evidence provided functional assignments or insights for 240 more genes (categories 2-5; 45%). These functional annotations advance our understanding of genes involved in vital cellular processes, including energy conversion, ion transport, secondary metabolism, and signal transduction. We propose that this integrative approach offers a valuable means to undertake the enormous challenge of characterizing the rapidly growing number of hypothetical proteins with each newly sequenced genome. computational biology | expression analysis | microarrays | proteomics | integrative microbiologyProceedings of The National Academy of Sciences - PNAS. 01/2005; 102(6):2099-2104. -
Article: LIP index for peptide classification using MS/MS and SEQUEST search via logistic regression.
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ABSTRACT: This study addresses the issue of peptide identification resulting from tandem mass spectrometry proteomics analysis followed by database search. This work shows that the Logistic Identification of Peptides (LIP) Index achieves high sensitivity and specificity for peptide classification relative to a manually verified "gold" standard and also accurately estimates the probability of a correct peptide match. The LIP Index is a weighted average of SEQUEST output variables based on logistic regression models and is a transparent, easy to use, inclusive, extendable, and statistically sound approach to classify correct peptide identifications. Modifications, such as normalizing cross-correlations (Xcorr) for peptide length, adjusting for charge state, and the number of tryptic termini, significantly improve the fit the logistic regression models, as well as increase sensitivity and specificity. The LIP Index also incorporates earlier developed statistical models on spectral quality assessment and peptide identification, which further improves sensitivity and specificity.Omics A Journal of Integrative Biology 02/2004; 8(4):357-69. · 2.44 Impact Factor -
Article: Spectral quality assessment for high-throughput tandem mass spectrometry proteomics.
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ABSTRACT: Current techniques in tandem mass spectrometric analyses of cellular protein contents often produce thousands to tens of thousands of spectra per experiment. This study introduces a new algorithm, named SPEQUAL, which is aimed at automated tandem mass spectral quality assessment. The quality of a given spectrum can be evaluated from three basic components: (i) charge state differentiation, (ii) total signal intensity, and (iii) signal-to-noise estimates. The differentiation between single and multiple precursor charge states (i) provides a binary score for a given spectrum. Components (ii) and (iii) provide partial scores which are subsequently summarized and multiplied by the first score. SPEQUAL was applied to over 10,000 data files derived from almost 3,000 tandem mass spectra, and the results (final cumulative scores) were manually verified. SPEQUAL's performance was determined to have high sensitivity and specificity and low error rates for both spectral quality estimates in general and precursor charge state differentiation in particular. Each of the partial scores is controlled by adjustable thresholds to fine-tune SPEQUAL's performance for different analysis pipelines and instrumentation. This spectral quality assessment tool is intended to act in an advisory role to the researcher, assisting in filtration of thousands of spectra typically produced by high throughput tandem mass spectrometric proteome analyses. Lastly, SPEQUAL was implemented as Java GUI-based and command-line-based interfaces freely available for both academic and industrial researchers.Omics A Journal of Integrative Biology 02/2004; 8(3):255-65. · 2.44 Impact Factor -
Article: Design and initial characterization of the SC-200 proteomics standard mixture.
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ABSTRACT: High-throughput (HTP) proteomics studies generate large amounts of data. Interpretation of these data requires effective approaches to distinguish noise from biological signal, particularly as instrument and computational capacity increase and studies become more complex. Resolving this issue requires validated and reproducible methods and models, which in turn requires complex experimental and computational standards. The absence of appropriate standards and data sets for validating experimental and computational workflows hinders the development of HTP proteomics methods. Most protein standards are simple mixtures of proteins or peptides, or undercharacterized reference standards in which the identity and concentration of the constituent proteins is unknown. The Seattle Children's 200 (SC-200) proposed proteomics standard mixture is the next step toward developing realistic, fully characterized HTP proteomics standards. The SC-200 exhibits a unique modular design to extend its functionality, and consists of 200 proteins of known identities and molar concentrations from 6 microbial genomes, distributed into 10 molar concentration tiers spanning a 1,000-fold range. We describe the SC-200's design, potential uses, and initial characterization. We identified 84% of SC-200 proteins with an LTQ-Orbitrap and 65% with an LTQ-Velos (false discovery rate = 1% for both). There were obvious trends in success rate, sequence coverage, and spectral counts with protein concentration; however, protein identification, sequence coverage, and spectral counts vary greatly within concentration levels.Omics: a journal of integrative biology 15(1-2):73-82. · 2.29 Impact Factor -
Article: Classifying proteins into functional groups based on all-versus-all BLAST of 10 million proteins.
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ABSTRACT: To address the monumental challenge of assigning function to millions of sequenced proteins, we completed the first of a kind all-versus-all sequence alignments using BLAST for 9.9 million proteins in the UniRef100 database. Microsoft Windows Azure produced over 3 billion filtered records in 6 days using 475 eight-core virtual machines. Protein classification into functional groups was then performed using Hive and custom jars implemented on top of Apache Hadoop utilizing the MapReduce paradigm. First, using the Clusters of Orthologous Genes (COG) database, a length normalized bit score (LNBS) was determined to be the best similarity measure for classification of proteins. LNBS achieved sensitivity and specificity of 98% each. Second, out of 5.1 million bacterial proteins, about two-thirds were assigned to significantly extended COG groups, encompassing 30 times more assigned proteins. Third, the remaining proteins were classified into protein functional groups using an innovative implementation of a single-linkage algorithm on an in-house Hadoop compute cluster. This implementation significantly reduces the run time for nonindexed queries and optimizes efficient clustering on a large scale. The performance was also verified on Amazon Elastic MapReduce. This clustering assigned nearly 2 million proteins to approximately half a million different functional groups. A similar approach was applied to classify 2.8 million eukaryotic sequences resulting in over 1 million proteins being assign to existing KOG groups and the remainder clustered into 100,000 functional groups.Omics: a journal of integrative biology 15(7-8):513-21. · 2.29 Impact Factor -
Article: 'Hypothetical' Genes and Improved Functional Annotations
Top co-authors
Top Journals
Institutions
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2010–2011
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Seattle Children’s Research Institute
Seattle, WA, USA
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2007–2011
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Seattle Children's Hospital
Seattle, WA, USA
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2004–2005
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University of Washington Bothell
Bothell, WA, USA
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