Abstract:
:Tyrosine phosphorylation plays a major role in regulating cell signaling pathways governing diverse biological functions such as proliferation and differentiation. Systemically mapping phosphotyrosine (pTyr) sites is the key to understanding molecular mechanisms underlining pTyr-dependent signaling. Although mass spectrometry-based technologies have been widely used for pTyr site profiling and quantification, their applications are often hindered by the poor efficiency in current multistep enrichment procedures for inherently low abundance pTyr peptides, especially under physiological conditions. Taking advantage of the sequence-independent high affinity of SH2 superbinder toward pTyr residues, we have developed a simplified one-step pTyr peptide enrichment method that uses immobilized SH2 superbinder for unbiased and robust enrichment of endogenous pTyr peptides from biological samples. By eliminating the prerequisite global phosphopeptide enrichment step in our previously developed two-step method, we minimized sample loss and improved peptide capture efficiency. Applying this method to Jurkat cells at resting state, where the tyrosine phosphorylation level is low, both the number of identified pTyr peptides and sites are increased by three folds compared to the two-step method. Specifically, we were able to identify 511 nonredundant pTyr peptides, corresponding to 403 high confidence pTyr sites, from Jurkat cells with high level technical reproducibility (Pearson's correlation coefficient as high as 0.94). Further applying this method to two human breast cancer cell lines, BT474 and HCC1954, before and after EGF stimulation, we demonstrated that this approach could be a powerful tool for illustrating pTyr-dependent signaling network controlling cellular behaviors such as drug resistance.
journal_name
J Proteome Resjournal_title
Journal of proteome researchauthors
Yao Y,Wang Y,Wang S,Liu X,Liu Z,Li Y,Fang Z,Mao J,Zheng Y,Ye Mdoi
10.1021/acs.jproteome.9b00045subject
Has Abstractpub_date
2019-04-05 00:00:00pages
1870-1879issue
4eissn
1535-3893issn
1535-3907journal_volume
18pub_type
杂志文章abstract::The study of glycosylation patterns (glycomics) in biological samples is an emerging field that can provide key insights into cell development and pathology. A current challenge in the field of glycomics is to determine how to quantify changes in glycan expression between different cells, tissues, or biological fluids...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr070476i
更新日期:2008-01-01 00:00:00
abstract::Phospholipase C gamma 1 (PLC-γ1) occupies a critically important position in the T-cell signaling pathway. While its functions as a regulator of both Ca2+ signaling and PKC-family kinases are well characterized, PLC-γ1's role in the regulation of early T-cell receptor signaling events is incompletely understood. Activ...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/acs.jproteome.6b01026
更新日期:2017-08-04 00:00:00
abstract::A new fast method for identification and characterization of proteolytic digests of proteins by monolithic liquid chromatography coupled with mass spectrometry has been developed. The advantages of the monolithic columns are a high-pressure stability and low back pressure resulting in higher flow rates for capillary o...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr060406w
更新日期:2007-02-01 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::Type-I procollagen aminoterminal propeptide (PINP) is a useful biomarker for bone formation activity that is used to monitor treatment of bone disorders including osteoporosis. Studies with human patients under long-term therapy for osteoporosis by daily injection of parathyroid hormone (PTH) demonstrated that the cir...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr070288s
更新日期:2007-11-01 00:00:00
abstract::Antibodies are a primary research tool for a diverse range of experiments in biology, from development to pathology. Their utility is derived from their ability to specifically identify proteins at a high level of sensitivity. This diversity of experimental requirements stretches the capabilities of these key research...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr100029f
更新日期:2010-07-02 00:00:00
abstract::Metabolomic approaches have the potential to make an exceptional contribution to understanding how chemicals and other environmental stressors can affect both human and environmental health. However, the application of metabolomics to environmental exposures, although getting underway, has not yet been extensively exp...
journal_title:Journal of proteome research
pub_type: 杂志文章,评审
doi:10.1021/pr060623x
更新日期:2007-02-01 00:00:00
abstract::We performed systematic evaluation of 38 protocols to concentrate normal human urinary proteins prior to 2D-PAGE analysis. Recovery yield and pattern of resolved protein spots were compared among different methods and intra-/inter-individual variabilities were examined. Precipitation with 90% ethanol provided the grea...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr0502525
更新日期:2006-01-01 00:00:00
abstract::The HIV-1-associated neurocognitive disorder occurs in approximately one-third of infected individuals. It has persisted in the current era of antiretroviral therapy, and its study is complicated by the lack of biomarkers for this condition. Since the cerebrospinal fluid is the most proximal biofluid to the site of pa...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr800854t
更新日期:2009-05-01 00:00:00
abstract::Quantitative comparison of protein expression levels in 2D gels is complicated by the variables associated with protein separation and mass spectrometric responses. Metabolic labeling allows cells from different experiments to be mixed prior to analysis. This approach has been reported for prokaryotic cells. Here, we ...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr025523f
更新日期:2002-07-01 00:00:00
abstract::The past decade has been marked by the emergence of selective affinity media and sensitive mass spectrometry instrumentation that facilitated large-scale phosphoproteome analyses and expanded the repertoire of protein phosphorylation. Despite these remarkable advances, the precise location of the phosphorylation site ...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr300229m
更新日期:2012-07-06 00:00:00
abstract::One of the critical challenges in predicting protein subcellular localization is how to deal with the case of multiple location sites. Unfortunately, so far, no efforts have been made in this regard except for the one focused on the proteins in budding yeast only. For most existing predictors, the multiple-site protei...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr060635i
更新日期:2007-05-01 00:00:00
abstract::S-Nitrosylation is a redox-based post-translational modification of a protein in response to nitric oxide (NO) signaling, and it participates in a variety of processes in diverse biological systems. The significance of this type of protein modification in health and diseases is increasingly recognized. In the central ...
journal_title:Journal of proteome research
pub_type: 杂志文章,评审
doi:10.1021/acs.jproteome.5b00857
更新日期:2016-01-04 00:00:00
abstract::The effects of acute and chronic stress on the production of systemic metabolites were investigated in male Sprague-Dawley (SD) rats. Metabolites excreted in urine were analyzed using GC/MS in conjunction with multivariate and univariate statistical techniques. SD rats were subjected to two kinds of acute stress and c...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr801086k
更新日期:2009-05-01 00:00:00
abstract::Protein oxidation and nitration have been described during spinal cord injury (SCI) in animal models. Herein, mass spectrometry unambiguously identified GDP-dissociation inhibitor-2 (GDI-2) in SCI with post-translational modifications of 3-aminotyrosine (8 h post-injury) and an acrolein adduct of GDI-2 (72 h post-inju...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr060620k
更新日期:2007-04-01 00:00:00
abstract::Mycobacterium tuberculosis, the causative agent of tuberculosis, accounts for 1.5 million human deaths annually worldwide. Despite efforts to eradicate tuberculosis, it still remains a deadly disease. The two best characterized strains of M. tuberculosis, virulent H37Rv and avirulent H37Ra, provide a unique platform t...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/acs.jproteome.6b00983
更新日期:2017-04-07 00:00:00
abstract::Reversible phosphorylations play a critical role in most biological pathways. Hence, in signaling studies great effort has been put into identification of a maximum number of phosphosites per experiment. Mass spectrometry (MS)-based phosphoproteomics approaches have been proven to be an ideal analytical method for map...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr200092z
更新日期:2011-08-05 00:00:00
abstract::For the 2018 YPIC Challenge, contestants were invited to try to decipher two unknown English questions encoded by a synthetic protein expressed in Escherichia coli. In addition to deciphering the sentence, contestants were asked to determine the three-dimensional structure and detect any post-translation modifications...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/acs.jproteome.9b00384
更新日期:2019-11-01 00:00:00
abstract::The majority of mitochondrial preproteins are targeted via N-terminal presequences that are cleaved upon import into the organelle. The essential mitochondrial processing protease (MPP) is assumed to cleave the majority of incoming precursors; however, only a small fraction of mitochondrial precursors have been experi...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/acs.jproteome.5b00327
更新日期:2015-11-06 00:00:00
abstract::The epidermal growth factor receptor (EGFR) is usually overexpressed in nasopharyngeal carcinoma (NPC) and is associated with pathogenesis of NPC. However, while EGFR-modulated intracellular proteins have been extensively studied, little is known concerning their extracellular counterparts. To identify EGFR-regulated ...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr100663p
更新日期:2010-12-03 00:00:00
abstract::Affinity purification coupled to 1-D gel-free liquid chromatography mass spectrometry (LC-MS) is a well-established and widespread approach for the analyses of noncovalently interacting protein complexes. In this study, two proteins conjugated to a streptavidin-binding peptide and hemagglutinin double tag were express...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr4009892
更新日期:2014-02-07 00:00:00
abstract::Plant secretory (Class III) peroxidases are redox enzymes that rely on N-glycosylation for full enzyme activity and stability. Peroxidases from palm tree leaves comprise the most stable and active plant peroxidases characterized to date. Herein, site-specific glycosylation and microheterogeneity of windmill palm tree ...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/acs.jproteome.6b00205
更新日期:2016-06-03 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::Ovarian granulosa cell tumors (GCTs) are hormonally active cancers characterized by indolent growth and late, invasive relapse. No therapies have yet proven to be efficacious. We previously reported that the inhibition of the antiapoptotic X-linked inhibitor of apoptosis protein (XIAP) removes transrepression of the p...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/acs.jproteome.8b00917
更新日期:2019-04-05 00:00:00
abstract::Proteoforms containing post-translational modifications (PTMs) represent a degree of functional diversity only harnessed through analytically precise simultaneous quantification of multiple PTMs. Here we present a method to accurately differentiate an unmodified peptide from its PTM-containing counterpart through data...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/acs.jproteome.0c00685
更新日期:2020-10-02 00:00:00
abstract::New technologies have advanced the field of proteomics, and a number of companies have developed innovative platforms to drive this research. However, significant challenges are often encountered when trying to integrate complementary technologies from multiple manufacturers. We have developed a software and hardware ...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr050465u
更新日期:2006-09-01 00:00:00
abstract::Human mesenchymal stromal cells (hMSCs) are promising candidates for cell therapy and tissue regeneration. Knowledge of the molecular mechanisms governing hMSC commitment into osteoblasts is critical to the development of therapeutic applications for human bone diseases. Because protein phosphorylation plays a critica...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr200868p
更新日期:2012-02-03 00:00:00
abstract::Bone proteomics is an expanding field for understanding protein changes associated with disease as well as characterizing and detecting proteins preserved in fossil bone. Most previous studies have utilized a protocol with demineralization and extraction approach to isolate and characterize proteins from bone. Through...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/acs.jproteome.7b00670
更新日期:2018-01-05 00:00:00
abstract::Interstitial cystitis/painful bladder syndrome (IC) is a chronic syndrome of unknown etiology that presents with bladder pain, urinary frequency, and urgency. The lack of specific biomarkers and a poor understanding of underlying molecular mechanisms present challenges for disease diagnosis and therapy. The goals of t...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr5007729
更新日期:2015-01-02 00:00:00
abstract::To investigate the role of rpoS in gene expression of Escherichia coli cells grown as biofilms, we compared the proteomes of a rpoS mutant and the wild-type strain. Experiments were performed on planktonic cells (in exponential or stationary growth phase) and biofilms developed on glass wool. Spot-by-spot comparison o...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr8001723
更新日期:2008-11-01 00:00:00