Phosphoprotein and phosphopeptide. Medical search. Frequent questions

With this present work, we report an approach to identify a large number of phosphopeptides and their phosphorylation sites using TiO2-centered phosphopeptide enrichment followed by LC-MS/MS analysis using high level of sensitivity nano-electrospray and an LTQ-Orbitrap instrument. (scienza-under-18.org)

The workflow is generally divided into two options: one for TiO 2 enrichment of phosphopeptides after the protein(s) of interest are purified (e.g. (yale.edu)
For the analysis of phosphoproteome, pooled and differentially labeled samples were initially mixed, digested to peptides and loaded into a Fe-IMAC column coupled to an HPLC system for the enrichment of phosphopeptides. (unifesp.br)

IMAC can be used to selectively enrich phosphopeptides from complex peptide mixtures, but co-retention of acidic peptides together with the failure to retain some phosphopeptides restricts the general utility of the method. (reading.ac.uk)

Among the identified phosphoproteins are flagellar motility proteins such as outer dynein arm, intraflagellar transport proteins as well as signaling molecules including protein kinases, phosphatases, G proteins, and ion channels. (nih.gov)
This analysis identified 9,643 phosphopeptides from 2,413 proteins, making it the "largest group of EV phosphoproteins [identified] to date," the authors wrote. (genomeweb.com)
The first is a more targeted approach to discover numerous sites of phosphorylation on proteins of interest, whereas the latter is directed towards a more global phosphopeptide discovery based approach. (yale.edu)
Sickle RBC treatment with U0126 decreased thirty-six phosphopeptides from twenty-one phosphoproteins involved in regulation of not only RBC shape, flexibility, cell morphology maintenance and adhesion, but also glucose and glutamate transport, cAMP production, degradation of misfolded proteins and receptor ubiquitination. (duke.edu)
To test this hypothesis, we compared the phosphopeptide-enriched fractions obtained from proteins secreted into conditioned media (CM) derived from five luminal and five basal type breast cancer cell lines using label-free quantitative mass spectrometry. (escholarship.org)
Proteins from cell lysate, uncooked serum, eluted serum proteins that were depleted of albumin and IgG, and eluted albumin and IgG from resin, were spiked with the phosphoprotein bovine beta-casein as an internal standard, reduced by 10 mM DTT in the presence of 8 M urea for 30 minutes at 37 C, and then alkylated by 50 mM iodoacetamide at space temp. (scienza-under-18.org)

Collectively, these results indicate the feasibility of using super-SILAC mix-SIM/AIMS assays for targeted verification of phosphopeptides discovered by large-scale phosphoproteome analyses of HCC specimens. (nih.gov)
This strategy allowed us to generate a quantitative HCC tissue phosphoproteome dataset containing significantly deregulated phosphoproteins that represents a valuable resource for the identification of potential HCC biomarkers and/or therapeutic targets. (nih.gov)
We want to identify and quantified phosphoproteins involved in manganese use efficiency, focusing on the phosphoproteome from thylakoid preparations from two barley genotypes, manganese efficient (Vanessa) and inefficient (Antonia) genotype. (sdu.dk)
Regarding the phosphoproteome experiment, 33 different phosphoproteins were identified, corresponding to 428 phosphopeptides. (unifesp.br)

Antibodies were purified by affinity-chromatography using epitope-specific phosphopeptide. (arigobio.cn)

Assessment of immunoaffinity (anti-phosphotyrosine antibody) versus chemical (TiO2) enrichment methods, as well as solitary versus dual enrichment strategies, yielded an initial assessment of the phosphoprotein/peptide content of human being serum, which can serve as a launch-point for further exploration and analysis. (scienza-under-18.org)
phosphopeptides ETTCSKEpSNEELTESCETK and AIPVAQDLNAPSDWDpSR were synthesized from Peptide 2.0 Inc.. The solitary clinical specimen chosen for analysis, a pre-operative, pre-treatment serum sample procured from a male subject with prostate malignancy undergoing radical prostatectomy, was acquired under IRB authorization (University or college of California-Irvine) and individual consent. (scienza-under-18.org)

The phosphopeptides were enriched by a combination of IMAC and titanium dioxide chromatography with a strategy of sequential elution from IMAC (SIMAC) and analyzed by tandem mass spectrometry. (nih.gov)

Eighty-nine of these phosphoproteins were only detected in shortening flagella, whereas 29 were solely in flagella of steady growing cells, indicating dramatic changes of protein phosphorylation during flagellar shortening. (nih.gov)
30,000 phosphopeptides in 12 h and protein-protein or protein-drug interaction experiments can be analyzed in 20 min per sample. (nature.com)
Vasodilator-stimulated phosphoprotein (VASP) is a member of the Ena-VASP protein family. (affbiotech.com)

This study utilized IMAC phosphoprotein isolation directly coupled with LC MS/MS analysis. (biomedcentral.com)

In this study Fe(III)-IMAC was qualitatively and quantitatively assessed using a panel of phosphopeptides, both synthetic and derived from proteolysis of known phosphoproteins, to identify the causes of success and failure in the application of this technique. (reading.ac.uk)
Modulating the loading buffer used for Fe(III)-IMAC significantly affects phosphopeptide binding and suggests that conformational factors that lead to steric hindrance and reduced accessibility to the phosphate are important. (reading.ac.uk)
The use of 1,1,1,3,3,3-hexa-fluoroisopropanol is shown here to significantly improve Fe(III)-IMAC enrichment and subsequent detection of phosphopeptides by MALDI-MS. (reading.ac.uk)

Sequencing of chymotryptic and thermolytic phosphopeptides from bovine DARPP-32 phosphorylated by casein kinase II suggested that the main phosphorylated residues were Ser45 and Ser102. (cornell.edu)

That said, the study suggests that EVs could prove a source of phosphoproteomics biomarkers and that it is possible to isolate and measure thousands of phosphoproteins from these structures. (genomeweb.com)
Our optimized TiO 2 phosphopeptide enrichment methodologies are combined with a state-of-the-art nanoACQUITY™ Ultra Performance or Ultra-high pressure LC (UPLC) system (Waters Corp.) /LTQ-Orbitrap XL mass spectrometer (Thermo Scientific) platform to provide a very robust phosphoprotein profiling and phosphoproteomics analysis. (yale.edu)
A multipathway phosphopeptide standard for rapid phosphoproteomics assay development. (stanford.edu)
Using mass spectrometry-based phosphoproteomics in data-independent acquisition allowed consistent quantification of over 3,000 phosphopeptides totaling more than 1,600 phosphoproteins across aposymbiotic (symbiont-free) and symbiotic anemones. (uni-konstanz.de)

Zirconium oxide is used by research scientists to isolate phosphopeptides. (bdlbiochem.com)

SDS PAGE) or immunoprecipitated and another for more complex phosphopeptide mixtures which are directly extracted from tissue or whole cell extracts. (yale.edu)

This mixture contains 131 mass spectrometry vetted phosphopeptides specifically chosen to cover as many known biologically interesting phosphosites as possible from seven different signaling networks: AMPK signaling, death and apoptosis signaling, ErbB signaling, insulin/IGF-1 signaling, mTOR signaling, PI3K/AKT signaling, and stress (p38/SAPK/JNK) signaling. (stanford.edu)

reported the profiling of endogenous serum phosphorylated peptides by TiO2-enrichment and MALDI-TOF MS detection.21 However, the characterization and description of the content of phosphoprotein analytes in serum were limited. (scienza-under-18.org)

We further demonstrate a comparison of phosphoproteomic profiling of HeLa performed independently in five labs using this phosphopeptide mixture with data-independent acquisition. (stanford.edu)

As a research group under the Association for Biomolecular Resource Facilities (ABRF) umbrella, the Proteomics Standards Research Group (sPRG) has worked to develop a multipathway phosphopeptide standard based on a mixture of heavy-labeled phosphopeptides designed to enable researchers to rapidly develop assays. (stanford.edu)

This analysis yielded 7868 quantifiable phosphopeptides and numerous up- or down-regulated candidates. (nih.gov)
Comparative analysis of phytohormone-responsive phosphoproteins in Arabidopsis thaliana using TiO2-phosphopeptide enrichment and mass accuracy precursor alignment. (mpg.de)
Recent advances in methodology have made phosphopeptide analysis a tractable problem for many proteomics researchers. (stanford.edu)
The analysis of phosphopeptides from breast cancer patient plasma and controls allowed us to construct a discovery list of phosphosites under rigorous collection conditions, and second to qualify discovery candidates generated from the CM studies. (escholarship.org)
This systems-level analysis defines the molecular framework in which EN oncofusions operate to promote cancer and provides some mechanisms for therapeutics. (bvsalud.org)

Altogether over 5000 phosphosites derived from 1756 phosphoproteins were identified, several of which have the potential to qualify as phosphopeptide plasma biomarker candidates for the more aggressive basal and also the luminal-type breast cancers. (escholarship.org)

Antibodies were produced by immunizing rabbits with KLH-conjugated synthetic phosphopeptide. (arigobio.cn)

To expand our verification capability, we evaluated the use of LTQ-Orbitrap run in SIM/Accurate inclusion mass screening (AIMS) mode with a super-SILAC mixture as an internal standard to quantify a subset of phosphopeptide candidates in HCC tissue samples. (nih.gov)
This method could be readily applied to verify dozens of phosphopeptide candidates in a larger HCC sample set. (nih.gov)
Together, this approach allowed us to assemble a set of novel cancer subtype specific phosphopeptide candidates for subsequent biomarker verification and clinical validation. (escholarship.org)

In this study, a phosphoproteomic approach was performed to dissect the molecular mechanism of phosphoprotein regulation, which is involved in the inhibitory effect of parasites on erythroid cell development. (biomedcentral.com)
Neuronal phosphoprotein that coats synaptic vesicles, binds to the cytoskeleton, and is believed to function in the regulation of neurotransmitter release. (arigobio.cn)

Our results suggest that widely available, biologically-relevant phosphopeptide standards can act as a quantitative "yardstick" across laboratories and sample preparations, enabling experimental designs larger than a single laboratory can perform. (stanford.edu)

In sample I used for discovery experiment, we successfully quantified 32 (in SIM mode) and 30 (in AIMS mode) phosphopeptides with median coefficients of variation (CVs) of 7.5% and 8.3%, respectively. (nih.gov)
When the assay was applied to other three pairs of HCC specimens for verification experiment, 40 target phosphopeptides were quantified reliably (~7.5% CV), and more than half of them were differentially expressed between tumor and adjacent non-tumor tissues. (nih.gov)

In a study published this week in the Proceedings of the National Academy of Sciences , the scientists identified 144 phosphoproteins in plasma EVs that were more highly phosphorylated in breast cancer patients compared to healthy controls. (genomeweb.com)

Comparison of thermolytic phosphopeptide maps and determination of the phosphorylated residue by manual Edman degradation identified the main phosphorylation site in intact cells as Ser102. (cornell.edu)

In this study, we developed a strategy for conducting both discovery and targeted verification of deregulated phosphoproteins in HCC tissue specimens on LTQ-Orbitrap. (nih.gov)
Furthermore, our proof-of-concept experiments demonstrated the feasibility of applying LTQ-Orbitrap, operated in SIM/AIMS mode, to multiplex and targeted verification of phosphopeptides in individual tissue specimens using a super-SILAC mix as an internal phosphopeptide standard. (nih.gov)

To provide molecular level insight into EN oncogenesis, we employed a proximity labeling mass spectrometry approach to define the molecular context of the fusions. (bvsalud.org)

The oncogenic ETV6-NTRK3 (EN) gene fusion joins the sterile alpha domain of the ETV6 transcription factor with the tyrosine kinase domain of the neurotrophin-3 receptor NTRK3. (bvsalud.org)

Here, we analyzed the phosphoproteins in the membrane/matrix fraction of flagella undergoing shortening as well as flagella from steady state cells of Chlamydomonas. (nih.gov)

We identify in total 237 high-confidence interactors, which link EN fusions to several key signaling pathways, including ERBB, insulin and JAK/STAT. We then assessed the effects of EN variants on these pathways, and showed that the pan NTRK inhibitor Selitrectinib (LOXO-195) inhibits the oncogenic activity of EN2, the most common variant. (bvsalud.org)

NEW YORK (GenomeWeb) - A team led by Purdue University researchers has identified a set of candidate breast cancer biomarkers by analyzing the phosphoprotein content of circulating extracellular vesicles (EVs). (genomeweb.com)

Using this approach we recognized ~100 unique phosphopeptides with stringent filtering criteria and a lower than 1% false discovery rate. (scienza-under-18.org)

The abundance of twelve of the thirty-six phosphopeptides were subsequently increased in normal RBCs co-incubated with recombinant ERK2 and therefore represent specific MEK1/2 phospho-inhibitory targets mediated via ERK2. (duke.edu)
This scholarly study highlights the and current limitations of the na?ve collection screening technique for phosphoprotein-specific antibody surrogates. (euromedh2020.com)
These phosphorylation state-specific antibodies (PSSAs) are nearly always made by immunization of pets with artificial phosphopeptides. (euromedh2020.com)
In addition, non-phospho specific antibodies were removed by chromatogramphy using non-phosphopeptide. (arigobio.cn)

Four EN variants with alternating break points have since been detected in a wide range of human cancers. (bvsalud.org)
The antiserum was produced against a chemically synthesized phosphopeptide derived from the region of human Shc that contains tyrosines 239 and 240. (thermofisher.com)

A total of 224 phosphoproteins derived from 1296 spectral counts of phosphopeptides were identified. (nih.gov)

The relative levels of forty four phosphoproteins were determined from gECs exposed to IE/UE for 24-72 h and compared with the media control using the label-free quantitation technique. (biomedcentral.com)

Anatomy25

Organisms23

Diseases1

Chemicals and Drugs137

Analytical, Diagnostic and Therapeutic Techniques and Equipment30

Phenomena and Processes40

Disciplines and Occupations1

Information Science3

Phosphorylation sites1

Enrichment of phosphopeptides2

Enrich phosphopeptides1

Proteins6

Phosphoproteome4

AFFINITY-CHROMATOGRAPHY1

Peptide2

Tandem mass spectro1

Protein3

Isolation1

IMAC3

Bovine1

Phosphoproteomics4

Isolate1

Mixtures1

Interesting phosphosites1

Characterization1

Data-independent acquisition1

Assays1

Analysis5

Basal1

Synthetic1

Candidates3

Regulation2

Quantitative1

Experiment2

Scientists1

Determination1

Verification2

Molecular1

Gene1

Fraction1

Identify1

Content1

Unique1

Specific4

Human2

Total1

Levels1