Abstract：
:The endothelium plays a critical role in orchestrating the inflammatory response seen during sepsis. Many of the inflammatory effects of Gram-negative sepsis are elicited by lipopolysaccharide (LPS), a glycolipid component of bacterial cell walls. Lipid-rich microdomains have been shown to concentrate components of the LPS signaling system. However, much remains to be learned about which proteins are constituents of lipid microdomains, and how these are regulated following cell activation. Progress in this area would be accelerated by employing global proteomic analyses, but the hydrophobicity of membrane proteins presents an analytical barrier to the effective application of such approaches. Herein, we describe a method to isolate detergent-resistant membranes from endothelial cells, and prepare these samples for proteomic analysis in a way that is compatible with subsequent separations and mass spectrometric (MS) analysis. In the application of these sample preparation and MS analyses, 358 proteins from the lipid-rich microdomains of LPS-activated endothelial cell membranes have been identified of which half are classified as membrane proteins by Gene Ontology. We also demonstrate that the sample preparation method used for solubilization and trypsin digestion of lipid-rich microdomains renders the membrane spanning sequences of transmembrane proteins accessible for endoproteolytic hydrolysis. This analysis sets the analytical foundation for an in-depth probing of LPS signaling in endothelial cells.
journal_name
J Proteome Resjournal_title
Journal of proteome researchauthors
Karsan A,Blonder J,Law J,Yaquian E,Lucas DA,Conrads TP,Veenstra Tdoi
10.1021/pr049824wkeywords:
subject
Has Abstractpub_date
2005-03-01 00:00:00pages
349-57issue
2eissn
1535-3893issn
1535-3907journal_volume
4pub_type
杂志文章abstract：:Tandem mass spectrometry (MS/MS) combined with bioinformatics tools have enabled fast and systematic protein identification based on peptide-to-spectrum matches. However, it remains challenging to obtain accurate identification of endogenous peptides, such as neuropeptides, peptide hormones, peptide pheromones, venom ...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr400446z
更新日期：2013-12-06 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：:Proteomics methods are being increasingly used to study archaeological and palaeontological bone, assisting in species identification and phylogenetic studies as well as improving our understanding of bone diagenesis. More recently, there are developing interests in the study of post-translational modifications, some ...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/acs.jproteome.6b00564
更新日期：2017-02-03 00:00:00
abstract：:Pathogenesis of Staphylococcus aureus, an opportunistic human pathogen, is complex and involves many virulence factors including an array of surface proteins (adhesins) that promote bacterial interactions with extracellular matrix components. A better understanding of these interactions can be achieved by studying the...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr049866k
更新日期：2005-03-01 00:00:00
abstract：:Serum protein profiling by mass spectrometry is a promising method for early detection of cancer. We have implemented a combined strategy based on matrix-assisted laser desorption ionization mass spectrometry (MALDI MS) and statistical data analysis for serum protein profiling and applied it in a well-described breast...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr7007576
更新日期：2008-04-01 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：:Advances in proteomics are continually driven by the introduction of new mass spectrometric instrumentation with improved performances. The recently introduced quadrupole Orbitrap (Q Exactive) tandem mass spectrometer allows fast acquisition of high-resolution higher-energy collisional dissociation (HCD) tandem mass s...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr3000249
更新日期：2012-06-01 00:00:00
abstract：:Glycosylation has significant effects on protein function and cell metastasis, which are important in cancer progression. It is of great interest to identify site-specific glycosylation in search of potential cancer biomarkers. However, the abundance of glycopeptides is low compared to that of nonglycopeptides after t...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/acs.jproteome.5b00068
更新日期：2015-04-03 00:00:00
abstract：:Our previous studies suggest that the α2,3sialylated T-antigen (NeuAcα2,3Galβ1,3GalNac-) and associated glycan structures are likely to be elevated during cancer. An easy and reliable strategy to label mucinous glycans that contain such carbohydrates can enable the identification of novel glycoproteins that are cancer...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr201108q
更新日期：2012-04-06 00:00:00
abstract：:Secreted proteins play a key role in cell signaling, communication, and migration. We recently described the development of an aggressive variant (T24M) of the bladder cancer cell line T24. Using this cell line model, the objective of our work was the identification of secreted proteins involved in the acquisition of ...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr100189d
更新日期：2010-06-04 00:00:00
abstract：:We report a hybrid search method combining database and spectral library searches that allows for a straightforward approach to characterizing the error rates from the combined data. Using these methods, we demonstrate significantly increased sensitivity and specificity in matching peptides to tandem mass spectra. The...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr101130b
更新日期：2011-05-06 00:00:00
abstract：:We undertook an unbiased metabolite profiling of fibroblasts from schizophrenia patients and healthy controls to identify metabolites and pathways that are dysregulated in disease, seeking to gain new insights into the disease biology of schizophrenia and to discover potential disease-related biomarkers. We measured p...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/acs.jproteome.6b00628
更新日期：2017-02-03 00:00:00
abstract：:Label-free quantification of shotgun proteomics data is a frequently used strategy, offering high dynamic range, sensitivity, and the ability to compare a high number of samples without additional labeling effort. Here, we present a bioinformatics approach that significantly improves label-free quantification results....
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/acs.jproteome.8b00113
更新日期：2019-01-04 00:00:00
abstract：:Lactic acid bacteria are broadly employed as starter cultures in the manufacture of foods. Upon technological preparation, they are confronted with drying stress that amalgamates numerous stress conditions resulting in losses of fitness and survival. To better understand and differentiate physiological stress response...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/acs.jproteome.7b00474
更新日期：2017-10-06 00:00:00
abstract：:Growing appreciation for astrocytes as active participants in nervous system development, neurovascular metabolic coupling, and neurological disease progression has stimulated recent investigation into specific astrocyte-secreted proteins that may mediate these functions. The current work utilized SILAC-generated isot...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr100134n
更新日期：2010-05-07 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：:Reported in this work are the development of mass spectrometric immunoassay (MSIA) devices and methods for the qualitative analysis of IGF-1 and -2, and the rigorous quantification of IGF-1 from human plasma. A method involving addition of SDS in moderate concentration to unfractionated plasma for disrupting IGF/IGFBP...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr0499388
更新日期：2004-07-01 00:00:00
abstract：:Proteomic studies were performed to identify the protein species involved in copper (Cu) stress responses in common wheat. Two-week-old wheat seedlings were exposed to 100 μM CuSO4 treatment for 3 days. Growth of shoots and roots was markedly inhibited and lipid peroxidation was greatly increased. Cu was readily absor...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr4008283
更新日期：2013-11-01 00:00:00
abstract：:Endothelial cells (ECs) form the inner layer of blood vessels and physically separate the blood from the surrounding tissue. To support tissues with nutrients and oxygen, the endothelial monolayer is semipermeable. When EC permeability is altered, blood vessels are not functional, and this is associated with disease. ...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/acs.jproteome.6b00166
更新日期：2016-07-01 00:00:00
abstract：:Although tandem mass spectrometry (MS/MS) has become an integral part of proteomics, intensity patterns in MS/MS spectra are rarely weighted heavily in most widely used algorithms because they are not yet fully understood. Here a knowledge mining approach is demonstrated to discover fragmentation intensity patterns an...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr070106u
更新日期：2008-01-01 00:00:00
abstract：:Selected reaction monitoring (SRM) is a technique that applies tandem mass spectrometry to quantify specific proteins of biological interest. The key to SRM is finding the best peptide-to-product ion transitions to monitor. The MRMaid database (MRMaid-DB) is a new online database for capturing SRM transitions from pub...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr900713u
更新日期：2010-01-01 00:00:00
abstract：:De Palma et al. published a research paper in which they describe the effect of chronic hypoxia on rat skeletal muscle metabolism by means of a comparative proteomic analysis (J. Proteome Res. 2007, 6(5), 1974-1984). For this, relatively young animals were used. In our communication, we note that, based on other liter...
journal_title：Journal of proteome research
pub_type： 评论,杂志文章
doi：10.1021/pr070315e
更新日期：2007-09-01 00:00:00
abstract：:Tomato fruit and seed allergens were detected by IgE-immunoblotting using sera from 18 adult tomato-sensitized patients selected based on a positive history skin prick test (SPT) and specific Immunglobulin (Ig) E-levels. Isolated tomato seed total protein showed high SPT activity comparable or even higher than tomato ...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr800186d
更新日期：2009-03-01 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：:Approximately 25% of eukaryotic proteins possessing homology to at least two transmembrane domains are predicted to be embedded in biological membranes. Nevertheless, this group of proteins is not usually well represented in proteome-wide experiments due to their refractory nature. Here we present a quantitative mass ...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr200759m
更新日期：2012-02-03 00:00:00
abstract：:Recombinant virus-like particles (VLPs) have been shown to induce protective immunity. Despite their potential significance as promising vaccine candidates, the protein composition of VLPs produced in insect cells has not been well characterized. Here we report a proteomic analysis of influenza VLPs containing hemaggl...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr200086v
更新日期：2011-08-05 00:00:00
abstract：:Neurodegenerative diseases are often defined pathologically by the presence of protein aggregates. These aggregates, including amyloid plaques in Alzheimer's disease (AD), result from the abnormal accumulation and processing of proteins, and may ultimately lead to neuronal dysfunction and cell death. To date, conventi...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr900474t
更新日期：2009-11-01 00:00:00
abstract：:Glioblastoma multiforme (GBM) is the most aggressive malignant primary brain tumor, with a dismal mean survival even with the current standard of care. Although in vitro cell systems can provide mechanistic insight into the regulatory networks governing GBM cell proliferation and migration, clinical samples provide a ...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr500418w
更新日期：2014-11-07 00:00:00
abstract：:Cyadox is an antibiotic drug and has the potential to be used as a feedstuff additive in promoting the growth of animals. However, the toxicity of cyadox should be fully assessed before application, and this has prompted the current investigation on the metabolic responses of mice to cyadox exposure, using a metabonom...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr301033a
更新日期：2013-01-04 00:00:00
abstract：:The notion of a gamete recognition system that alerts females to the presence of gametes in their reproductive tract profoundly influences our understanding of the physiology of events leading to conception and the bearing of offspring. Here, we show that the female responds to gametes within her tract by modulating t...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr070349m
更新日期：2007-12-01 00:00:00
