Abstract：
:Genomic-scale analysis of protein function is currently limited by the ability to rapidly express the enormous diversity of protein targets in their active form. We describe a method to construct transcriptionally active expression templates (ETs) in parallel using a single PCR step wherein the overlap-extension reaction for addition of transcription regulatory elements is separated from the amplification of the full-length product by using a GC-rich single primer. Over 90% of 55 diverse genomic targets were extended with T7 regulatory elements to form ETs in high yield and purity. The unpurified ETs directed protein expression using a cell-free protein synthesis (CFPS) system supplemented with cofactors and metal ions to activate a variety of enzymes. Higher activities were obtained in the modified CFPS reactions compared to standard reaction conditions. Protein purification was avoided because the expressed enzyme activity was significantly greater than the background activity associated with the cell extract. These improvements in the parallel synthesis of linear ETs combined with enhanced in vitro enzyme activation help to make CFPS systems more attractive platforms for high-throughput evaluation of protein function.
journal_name
J Proteome Resjournal_title
Journal of proteome researchauthors
Woodrow KA,Airen IO,Swartz JRdoi
10.1021/pr050459ysubject
Has Abstractpub_date
2006-12-01 00:00:00pages
3288-300issue
12eissn
1535-3893issn
1535-3907journal_volume
5pub_type
杂志文章abstract：:A metabonomic study was performed to investigate the biochemical perturbation of the serum samples from liver failure patients induced by hepatitis B virus (HBV; n=24) and control normal subjects (n=23). The serum metabonome was detected using gas chromatography-mass spectrometry (GC-MS) technique integrated with a co...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr060591d
更新日期：2007-07-01 00:00:00
abstract：:Whole cell protein and outer membrane protein (OMP) extracts were compared for their ability to differentiate and delineate the correct database organism to an experimental sample and for the degree of dissimilarity to the nearest neighbor database organism strains. These extracts were isolated from pathogenic and non...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr100402y
更新日期：2010-07-02 00:00:00
abstract：:The urine metabotype of 12 individuals was followed over a period of 8-10 years, which provided the longest longitudinal study of metabolic phenotypes to date. More than 2000 NMR metabolic profiles were analyzed. The majority of subjects have a stable metabotype. Subjects who were exposed to important pathophysiologic...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/acs.jproteome.5b00275
更新日期：2015-07-02 00:00:00
abstract：:We investigated the effect of homogenization strategy and protein precipitation on downstream protein quantitation using multiple reaction monitoring mass spectrometry (MRM-MS). Our objective was to develop a workflow capable of processing disparate tissue types with high throughput, minimal variability, and maximum p...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/acs.jproteome.0c00399
更新日期：2021-01-01 00:00:00
abstract：:Knowledge of protein subcellular localization assists in the elucidation of protein function and understanding of different biological mechanisms that occur at discrete subcellular niches. Organelle-centric proteomics enables localization of thousands of proteins simultaneously. Although such techniques have successfu...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr4008464
更新日期：2014-02-07 00:00:00
abstract：:Pancreas ductal adenocarcinoma (PDAC) remains a deadly malignancy with poor early diagnostic and no effective therapy. Although several proteomic studies have performed comparative analysis between normal and malignant tissues, there is a lack of clear characterization of proteins that could be of clinical value. Syst...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr200527z
更新日期：2011-09-02 00:00:00
abstract：:Large-scale phosphoproteomic analysis employing liquid chromatography-tandem mass spectrometry (LC-MS/MS) often requires a significant amount of manual manipulation of phosphopeptide datasets in the post-acquisition phase. To assist in this process, we have created software, PhosphoPIC (PhosphoPeptide Identification a...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr0701153
更新日期：2007-09-01 00:00:00
abstract：:Hepatitis C virus (HCV) causes chronic liver disease worldwide. HCV Core protein (Core) forms the viral capsid and is crucial for HCV pathogenesis and HCV-induced hepatocellular carcinoma, through its interaction with the host factor proteasome activator PA28γ. Here, using BD-PowerBlot high-throughput Western array, w...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr300121a
更新日期：2012-07-06 00:00:00
abstract：:Ovarian cancer remains a deadly threat to women as the disease is often diagnosed in the late stages when the chance of survival is low. There are no good biomarkers available for early detection and only a few markers have shown clinical utility for prognosis, response to therapy and disease recurrence. We mined cond...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr900411g
更新日期：2009-10-01 00:00:00
abstract：:Many proteins possess intrinsic disorder (ID) and lack a rigid three-dimensional structure in at least part of their sequence. ID has been hypothesized to influence protein-protein and protein-ligand interactions. We calculated ID for nearly 400 vertebrate and invertebrate members of the biomedically important nuclear...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr8003024
更新日期：2008-10-01 00:00:00
abstract：:Compartment proteomics enable broad characterization of target tissues. We employed a simple fractionation method and filter-aided sample preparation (FASP) to characterize the cytosolic and membrane fractions of white perch ovary tissues by semiquantitative tandem mass spectrometry using label-free quantitation based...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr401067g
更新日期：2014-03-07 00:00:00
abstract：:Difference gel electrophoresis (DiGE) allows the reliable comparison of proteome differences between two or three samples within a single gel, by way of a CyDye fluorescent labeling system. This facilitates identification of protein differences avoiding the difficulties associated with gel-to-gel variation. A drawback...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr0606878
更新日期：2007-09-01 00:00:00
abstract：:Bacterial small RNAs (sRNAs) and two-component systems (TCSs) were two vital regulatory mechanisms employed by microorganisms to respond to environmental changes and stresses. As a promising "autotrophic cell factory", photosynthetic cyanobacteria have attracted a lot of attention these years. Although most studies fo...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/acs.jproteome.7b00243
更新日期：2017-08-04 00:00:00
abstract：:In humans, the olfactory epithelium is located in two narrow passages, the olfactory clefts, at the upper part of the nasal cavities. The olfactory epithelium is covered by a mucus layer which is essential for the function of the olfactory neurons that are directly connected with the brain through the cribriform plate...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr0606575
更新日期：2007-05-01 00:00:00
abstract：:The complete and specific proteolytic cleavage of protein samples into peptides is crucial for the success of every shotgun LC-MS/MS experiment. In particular, popular peptide-based label-free and targeted mass spectrometry approaches rely on efficient generation of fully cleaved peptides to ensure accurate and sensit...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr300273g
更新日期：2012-11-02 00:00:00
abstract：:Papillary thyroid carcinoma (PTC) is the most common type of thyroid cancer among women worldwide. It is confirmed mainly by fine-needle aspiration biopsy (FNAB), an invasive diagnostic method. The key proteins responsible for thyroid hormone biosynthesis are glycosylated. Hence, changes in site-specific glycosylation...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/acs.jproteome.9b00672
更新日期：2020-07-02 00:00:00
abstract：:A synthetic coumarin, N,N,N-triethyl-11-(4-methyl-2-oxo-2H-chromen-7-yloxy)-11-oxoundecan-1-aminium bromide (SCD-1), having potent activity against pathogenic Aspergilli (MIC90 15.62 μg/mL), was investigated to identify its molecular targets in the pathogen. The proteome of Aspergillus fumigatus was developed after tr...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr300006j
更新日期：2012-06-01 00:00:00
abstract：:In an attempt to find out the association of metabolic dysregulation with poor endometrial receptivity and pregnancy loss, serum metabonomic profiling of women with idiopathic recurrent spontaneous miscarriage (IRSM) is carried out and compared with fertile controls. (1)H nuclear magnetic resonance (NMR)-based metabon...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr500379n
更新日期：2014-06-06 00:00:00
abstract：:Lung cancer, with its high metastatic potential and high mortality rate, is the worldwide leading cause of cancer-related deaths. High-throughput "omics"-based platforms have accelerated the discovery of biomarkers for lung cancer, and the resulting candidates are to be evaluated for their diagnostic potential as noni...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr5010828
更新日期：2015-03-06 00:00:00
abstract：:The final goal of the Russian part of the Chromosome-centric Human Proteome Project (C-HPP) was established as the analysis of the chromosome 18 (Chr 18) protein complement in plasma, liver tissue and HepG2 cells with the sensitivity of 10(-18) M. Using SRM, we have recently targeted 277 Chr 18 proteins in plasma, liv...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr300821n
更新日期：2013-01-04 00:00:00
abstract：:Oxidative stress and inflammation play a role in the physiopathology of insulin resistance, diabetes and cardiovascular disease. A single high-fat, high-carbohydrate (HFHC) meal induces an increase in inflammatory and oxidative stress markers in peripheral blood mononuclear cells (PBMC). Previous studies have shown th...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/acs.jproteome.7b00476
更新日期：2017-11-03 00:00:00
abstract：:Advances in liquid chromatography tandem mass spectrometry (LC-MS/MS) have permitted phosphoproteomic analysis on a grand scale, but ongoing challenges specifically associated with confident phosphate localization continue to motivate the development of new fragmentation techniques. In the present study, ultraviolet p...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/acs.jproteome.6b00289
更新日期：2016-08-05 00:00:00
abstract：:Staphylococcus aureus with spa-type t437 has been identified as a predominant community-associated methicillin-resistant S. aureus clone from Asia, which is also encountered in Europe. Molecular typing has previously shown that t437 isolates are highly similar regardless of geographical regions or host environments. T...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/acs.jproteome.9b00179
更新日期：2019-07-05 00:00:00
abstract：:Complicating proteomic analysis of whole tissues is the obvious problem of cell heterogeneity in tissues, which often results in misleading or confusing molecular findings. Thus, the coupling of tissue microdissection for tumor cell enrichment with capillary isotachophoresis-based selective analyte concentration not o...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr300303t
更新日期：2012-08-03 00:00:00
abstract：:beta-Cell granules contain proteins involved in fuel regulation, which when altered, contribute to metabolic disorders including diabetes mellitus. We analyzed proteins present in purified granules from the INS-1E beta-cell model. Fifty-one component proteins were identified by LC-MS/MS including hormones, granins, pr...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr800675k
更新日期：2009-01-01 00:00:00
abstract：:The paper describes the implementation of a software system based on the Fenyö disulfide bond assignment algorithm. The system allows an investigator to enter data derived from mass spectrum peak assignments, a target protein sequence and other experimental conditions. The output of the system is the set of disulfide ...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr034016a
更新日期：2003-11-01 00:00:00
abstract：:During vertebrate central nervous system development, the apical neuroepithelium is bathed with embryonic Cerebrospinal Fluid (e-CSF) which plays regulatory roles in cortical cell proliferation and maintenance. Here, we report the first proteomic analysis of human e-CSF and compare it to an extensive proteomic analysi...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr070247w
更新日期：2007-09-01 00:00:00
abstract：:Bacillus Amyloliquefaciens SQR9 is a well-investigated plant growth-promoting rhizobacteria with strong root colonization capability. To identify the key proteins involved in in situ root colonization and biofilm formation, the proteomic profiles of planktonic and root colonized SQR9 cells were compared. A total of 75...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr500565m
更新日期：2014-12-05 00:00:00
abstract：:The purpose of this study was to evaluate the use of high resolution LC-MS together with metabolomics and D(4)-cholic acid (D(4)-CA) as a metabolic tracer to measure the metabolism and reconjugation of bile acids (BAs) in vitro and in vivo. Metabolic tracers are very important because they allow for the direct detecti...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr200475g
更新日期：2011-10-07 00:00:00
abstract：:Inspired by the availability of de novo transcriptome of horse gram (Macrotyloma uniflorum) and recent developments in systems biology studies, the first ever global protein-protein interactome (PPI) map was constructed for this highly drought-tolerant legume. Large-scale studies of PPIs and the constructed database w...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/acs.jproteome.5b01114
更新日期：2016-06-03 00:00:00
