Abstract：
:The search for novel enzymes is an important but difficult task in functional genomics. Here, we present a systematic method based on in vitro assays in combination with metabolite profiling to discover novel enzymatic activities. A complex mixture of metabolites is incubated with purified candidate proteins and the reaction mixture is subsequently profiled by capillary electrophoresis electrospray ionization mass spectrometry (CE-MS). Specific changes in the metabolite composition can directly suggest the presence of an enzymatic activity while subsequent identification of the compounds whose level changed specifically can pinpoint the actual substrate(s) and product(s) of the reaction. We first evaluated the method using several Escherichia coli metabolic enzymes and then applied it to the functional screening of uncharacterized proteins. In this manner, YbhA and YbiV proteins were found to display both phosphotransferase and phosphatase activity toward different sugars/sugar phosphates. Our approach should be broadly applicable and useful for enzyme discovery in any system.
journal_name
J Proteome Resjournal_title
Journal of proteome researchauthors
Saito N,Robert M,Kitamura S,Baran R,Soga T,Mori H,Nishioka T,Tomita Mdoi
10.1021/pr0600576subject
Has Abstractpub_date
2006-08-01 00:00:00pages
1979-87issue
8eissn
1535-3893issn
1535-3907journal_volume
5pub_type
杂志文章abstract：:In this paper, we describe a method for analyzing large-volume protein samples using capillary electrophoresis in conjunction with laser-induced fluorescence detection (CE-LIF). To improve the stacking and separation efficiencies of proteins, we added either 0.01% sodium dodecyl sulfate (SDS) or 0.01% poly(ethylene ox...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr050298y
更新日期：2006-02-01 00:00:00
abstract：:Metal based chemotherapeutic drugs are widely used as an effective method to defeat various cancers. In this study, the mechanism of action of a novel therapeutic agent, [Pd(sac)(terpy)](sac)·4H2O (sac = saccharinate, and terpy = 2,2':6',2″-terpyridine) was studied. To better understand the proteomic changes in respon...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr5006718
更新日期：2014-11-07 00:00:00
abstract：:A proteome of a model organism, Caenorhabditis elegans, was analyzed by an integrated liquid chromatography (LC)-based protein identification system, which was constructed by microscale two-dimensional liquid chromatography (2DLC) coupled with electrospray ionization (ESI) tandem mass spectrometry (MS/MS) on a high-re...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr025551y
更新日期：2003-01-01 00:00:00
abstract：:12-O-Tetradecanoyl-phorbol-13-acetate (TPA) is a plant derivative with multiple function as tumor promoter, differentiation revulsant or leukemia therapy drug. The molecular mechanism of its function is perplexing. Many studies have focused on the mechanism of TPA stimulation in tumor promotion of mouse models or term...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr0497677
更新日期：2005-03-01 00:00:00
abstract：:Two-dimensional differential gel electrophoresis (2-D DIGE) was used to analyze human serum following the removal of albumin and five other high-abundant serum proteins. After protein removal, serum was analyzed by SDS-PAGE as a preliminary screen, and significant differences between four high-abundant protein removal...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr049921p
更新日期：2004-11-01 00:00:00
abstract：:T lymphocytes mediate cellular and humoral defense against foreign bodies or autoantigens. An understanding of T-cell information processing furthers studies of the immunological response. We describe a large-scale phosphorylation analysis of primary T cells using a multidimensional separation strategy, involving prep...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr800500r
更新日期：2008-12-01 00:00:00
abstract：:To assess the global effect of preterm birth on fetal metabolism and maternal-fetal nutrient transfer, we used a mass spectrometric-based chemical phenotyping approach on cord blood obtained at the time of birth. We sampled umbilical venous, umbilical arterial, and maternal blood from mothers delivering very-low birth...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr400122v
更新日期：2013-06-07 00:00:00
abstract：:To gain insights into how TGF-beta regulates epithelial-mesenchymal transition (EMT), we assessed the time course of proteins and mRNAs during EMT by multiplex iTRAQ labeling and 2D-LC-MS/MS, and by hybridization, respectively. Temporal iTRAQ analysis identified 66 proteins as differentially expressed during EMT, incl...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr8006478
更新日期：2009-01-01 00:00:00
abstract：:High-resolution proton nuclear magnetic resonance (NMR) spectrometry of biofluids has been increasingly used in laboratory diagnosis of various diseases. In this study, we extended the use of (1)H NMR spectroscopy for laboratory diagnosis of exudative pleural effusions using pleural fluids. We compared this new NMR-ba...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr5004856
更新日期：2014-09-05 00:00:00
abstract：:Glycolysis is the core metabolic pathway supplying energy to cells. Whereas the vast majority of studies focus on specific aspects of the process, global analyses characterizing simultaneously all enzymes involved in the process are scarce. Here, we demonstrate that quantitative label- and standard-free proteomics all...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/acs.jproteome.5b00276
更新日期：2015-08-07 00:00:00
abstract：:In this Letter, we reanalyze published mass spectrometry data sets of clinical samples with a focus on determining the coinfection status of individuals infected with SARS-CoV-2 coronavirus. We demonstrate the use of ComPIL 2.0 software along with a metaproteomics workflow within the Galaxy platform to detect cohabita...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/acs.jproteome.0c00822
更新日期：2021-01-04 00:00:00
abstract：:We report the use of chemical derivatization with MALDI-MS/MS analysis for de novo sequence analysis. Using three frequently used homology-based search algorithms, we were able to identify more than 40 proteins from banana, a nonmodel plant with unsequenced genome. Furthermore, this approach allowed the identification...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr0601943
更新日期：2007-01-01 00:00:00
abstract：:The proteome of any system is a dynamic entity, such that the intracellular concentration of a protein is dictated by the relative rates of synthesis and degradation. In this work, we have analyzed time-dependent changes in the incorporation of a stable amino acid resolved precursor, a protocol we refer to as "dynamic...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr800641v
更新日期：2009-01-01 00:00:00
abstract：:Reliable statistical validation of peptide and protein identifications is a top priority in large-scale mass spectrometry based proteomics. PeptideProphet is one of the computational tools commonly used for assessing the statistical confidence in peptide assignments to tandem mass spectra obtained using database searc...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr7006818
更新日期：2008-01-01 00:00:00
abstract：:Frontotemporal lobar degeneration (FTLD) is an umbrella term for a heterogeneous group of young-onset dementias of uncertain prevalence and incidence worldwide. Atypical cases of FTLD with fused in sarcoma inclusions (aFTLD-U) have been described recently, but their molecular characterization is still due. Using shotg...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr2012279
更新日期：2012-04-06 00:00:00
abstract：:Rosiglitazone is a thiazolidinedione used to treat insulin resistance in diabetes. Although thiazolidinediones may also exert cardiovascular effects, contrasting results were reported. Favorable effects were shown for pioglitazone, whereas adverse reactions were suspected for rosiglitazone. Therefore, a reassessment o...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr900435z
更新日期：2010-01-01 00:00:00
abstract：:Automated sequencing of unknowns in bottom-up proteomics makes the data produced susceptible to process control errors, which can be propagated into mistakes in analyte identification. Inclusion of an unintrusive internal standard, such as lysozyme, allows monitoring all phases of the proteomics process including samp...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr049819s
更新日期：2005-01-01 00:00:00
abstract：:While the proteome defines the expressed gene products, the metabolome results from reactions controlled by such gene products. Plasma represents an accessible "window" to the metabolome both in regard of availability and content. The wide range of the plasma metabolome, in terms of molecular diversity and abundance, ...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr8001639
更新日期：2008-08-01 00:00:00
abstract：:Flooding is a serious problem for soybeans because it reduces growth and grain yield. Proteomic and metabolomic techniques were used to examine whether mitochondrial function is altered in soybeans by flooding stress. Mitochondrial fractions were purified from the roots and hypocotyls of 4-day-old soybean seedlings th...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr2001918
更新日期：2011-09-02 00:00:00
abstract：:Cannabinoid 2 receptor (CB2R), a Class-A G-protein coupled receptor (GPCR), is a promising drug target under a wide array of pathological conditions. Rational drug design has been hindered due to our poor understanding of the structural features involved in ligand binding. Binding of a high-affinity biarylpyrazole inv...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/acs.jproteome.7b00023
更新日期：2017-07-07 00:00:00
abstract：:Many pressing medical challenges, such as diagnosing disease, enhancing directed stem-cell differentiation, and classifying cancers, have long been hindered by limitations in our ability to quantify proteins in single cells. Mass spectrometry (MS) is poised to transcend these limitations by developing powerful methods...
journal_title：Journal of proteome research
pub_type： 杂志文章,评审
doi：10.1021/acs.jproteome.8b00257
更新日期：2018-08-03 00:00:00
abstract：:We report the structural analysis of cap-binding proteins using a chemical probe/ultraviolet photodissociation (UVPD) mass spectrometry strategy for evaluating solvent accessibility of proteins. Our methodology utilized a chromogenic probe (NN) to probe the exposed amine residues of wheat eukaryotic translation initia...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr400869u
更新日期：2013-12-06 00:00:00
abstract：:Epithelial cells play an important role in physiological and pathophysiological situations, with organ-, tissue-, type-, and function-specific patterns. Proteome analysis has been used to study epithelial-origin diseases and identify novel prognostic, diagnostic, and therapeutic markers. The present review compares th...
journal_title：Journal of proteome research
pub_type： 杂志文章,评审
doi：10.1021/pr050389v
更新日期：2006-04-01 00:00:00
abstract：:In an attempt to identify prostate cancer biomarkers with greater diagnostic and prognostic capabilities, we have developed an integrative proteomic discovery workflow focused on N-linked glycoproteins that refines the target selection process. In this work, hydrazide-based chemistry was used to identify N-linked glyc...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr201200n
更新日期：2012-05-04 00:00:00
abstract：:Outbreaks of porcine epidemic diarrhea virus (PEDV) have caused significant lethality rates in neonatal piglets, which pose a serious threat to the swine industry worldwide. Available commercial vaccines fail to protect against the emergence of high virulence of PEDV variants. Therefore, the endemic state of the PEDV ...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/acs.jproteome.8b00897
更新日期：2019-04-05 00:00:00
abstract：:The balance between comprehensively analyzing the proteome and using valuable mass spectrometry time is a genuine challenge in the field of proteomics. Multidimensional fractionation strategies have significantly increased proteome coverage, but often at the cost of increased mass analysis time, despite advances in ma...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/acs.jproteome.9b00082
更新日期：2019-05-03 00:00:00
abstract：:Cellular functions are largely carried out by noncovalent protein complexes that may exist within the cell as stable modules or as assemblies of dynamically changing composition, whose formation and decomposition are triggered in response to extracellular stimuli. The protein constituents of complexes often exhibit po...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr1003888
更新日期：2010-10-01 00:00:00
abstract：:Ozonated autohemotherapy (O3-AHT) is a medical approach during which blood obtained from the patient is ozonated and injected back into the body. Despite an increasing number of evidence that O3-AHT is safe, this type of therapy remains controversial. To extend knowledge about the changes in blood evoked by O3-AHT, LC...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr3008946
更新日期：2012-12-07 00:00:00
abstract：:The peptide toxins in the venoms of small invertebrates such as stinging ants have rarely been studied due to the limited amount of venom available per individual. We used a venomics strategy to identify the molecular diversity of the venom peptidome for the myrmicine ant Tetramorium bicarinatum. The methodology inclu...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/acs.jproteome.8b00452
更新日期：2018-10-05 00:00:00
abstract：:Expansion of human articular chondrocytes (HACs) in serum-free medium (SFM) has been shown to be enhanced by the control of medium osmolality. HACs attained better growth when cultured at osmolalities lower than the average osmolality found in articular cartilage in vivo, although the specific causes for this improved...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr100136q
更新日期：2010-05-07 00:00:00
