Phosphorylation phosphopeptides. Medical search. Frequent questions

This study aimed to generate additional information on TTP phosphorylation using phosphopeptide mapping and mass spectrometry (MS). Wild-type and site-directed mutant TTP proteins were expressed in transfected human cells followed by in vivo radiolabeling with [32P]-orthophosphate. (usda.gov)
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)
Monoclonal antibodies PAb246 and PAb240 were used to immunoprecipitate metabolically 32P-labelled p53 protein in the 'wild-type' or mutant conformation from cells grown at 32 degrees C or 37 degrees C. Tryptic phosphopeptide maps were prepared from the isolated 'wild-type' and mutant p53 proteins. (dundee.ac.uk)
In a targeted analysis measuring phosphorylation levels of four of these 144 proteins, they found in three of the four - RALGAPA2, PKG1, and TJP2 - significant differences between levels in breast cancer patients and controls. (genomeweb.com)
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)
From this set, they identified 144 proteins whose phosphorylation levels differed significantly between cases and controls, and, using parallel-reaction monitoring mass spec analysis on a Thermo Fisher Q Exactive HF, they confirmed these observed differences in the aforementioned three proteins. (genomeweb.com)
As one of the most common cellular signaling processes, protein phosphorylation has been a large focus of cancer biomarker research, and kinases, the proteins responsible for protein phosphorylation, are a significant and growing area in drug development. (genomeweb.com)
Flow chart of the FACE in cell Western method that uses a cell based ELISA to measure the levels of the native and phospho forms of signaling proteins and kinases that are activated by phosphorylation. (activemotif.com)
These findings suggest that parasite proteins are able to inhibit erythroid cell growth by down-regulation of ezrin phosphorylation, leading to ineffective erythropoiesis ultimately resulting in severe malarial anaemia. (biomedcentral.com)
Many molecular pathways in eukaryotic cells are modulated by specific signaling proteins that are controlled, by phosphorylation and dephosphorylation, through the activity of kinase and phosphatase enzymes. (biomedcentral.com)
Using heavy isotope variants of individual phosphopeptides enables quantification by mass spectrometry of key proteins in signal transduction, such as in cancer research studying new kinase inhibitors. (panatecs.com)
A group of enzymes that catalyzes the phosphorylation of serine or threonine residues in proteins, with ATP or other nucleotides as phosphate donors. (lookformedical.com)

Liu Y, Zhang S (2004) Phosphorylation of 1-aminocyclopropane-1-carboxylic acid synthase by MPK6, a stress-responsive mitogen-activated protein kinase, induces ethylene biosynthesis in Arabidopsis . (springer.com)
The phosphorylation of IκBα by IκB kinase family have been so many identified, but the phosphorylation sites of IκBα by other kinases remain poorly understood. (researchsquare.com)
To identify the essential domains of IκBα for phosphorylation of IκBα by AURK, kinase assay was performed with a series of IκBα truncation mutants. (researchsquare.com)
Stable isotope labeling of phosphopeptides for multiparallel kinase target analysis and identification of phosphorylation sites. (mpg.de)
1998) Regulation of the p70 S6 kinase by phosphorylation in vivo. (concordia.ca)
RESULTS: To further characterize global ERK1/2-induced changes in membrane protein phosphorylation within human RBCs, a label-free quantitative phosphoproteomic analysis was applied to sickle and normal RBC membrane ghosts pre-treated with U0126, a specific inhibitor of MEK1/2, the upstream kinase of ERK1/2, in the presence or absence of recombinant active ERK2. (duke.edu)
Gene therapy displacement of endogenous MCUb with a dominant-negative MCUb transgene (MCUb W246R/V251E ) in vivo rescued T2D cardiomyocytes from metabolic inflexibility and stimulated cardiac contractile function and adrenergic responsiveness by enhancing phospholamban phosphorylation via protein kinase A. We conclude that MCUb represents one newly discovered molecular effector at the interface of metabolism and cardiac function, and its repression improves the outcome of the chronically stressed diabetic heart. (diabetesjournals.org)
TLK16998 alone had no effect on IR signaling in mouse 3T3-L1 adipocytes but, at concentrations as low as 3.2 μmol/l, enhanced the effects of insulin on the phosphorylation of the IR β-subunit and IR substrate 1, and on the amount of phosphatidylinositol 3-kinase that coimmunoprecipitated with IRS-1. (diabetesjournals.org)
Phosphopeptide mapping revealed that the effect of TLK16998 on the IR was associated with increased tyrosine phosphorylation of the activation loop of the β-subunit tyrosine kinase domain. (diabetesjournals.org)
Each response was separated using SDS-Web page, stained with Coomassie and the band corresponding to Raptor was excised, trypsin digested, enriched for phosphopeptides and then analyzed by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). The peptides identified from the second reaction are presumed to be from another kinase that could be co-purified from cells with HA-Raptor, such as mTOR. (healthweblognews.info)

Phosphorylation of serine and threonine residues is conserved during MHC class I and MHC class II antigen processing. (wikipedia.org)
Finally, by combining the sequence recognition model with the quantitative changes in phosphorylation measured in the two drug perturbation experiments, we identified 29 reliable candidates of PKA targeting residues in living cells including 8 previously known substrates. (nih.gov)
In mouse hepatocytes, p38γ induces proliferation after partial hepatectomy by promoting the phosphorylation of retinoblastoma tumour suppressor protein at known CDK target residues. (nature.com)
PDPK1 activates conventional PKC and PKC zeta through phosphorylation of critical threonine residues in the activation loop. (thermofisher.com)
Phosphorylation of serine residues in the N-terminus modulates the activity of ACA8, a plasma membrane Ca2+-ATPase of Arabidopsis thaliana / S. Giacometti, C.A. Marrano, M.C. Bonza, L. Luoni, M. Limonta, M.I. De Michelis. (unimi.it)

KLH conjugated synthetic phosphopeptide derived from human PDPK1 around the phosphorylation site of Ser393. (thermofisher.com)

Plentiful studies have established a close association between aberrant phosphorylation and hepatocellular carcinoma (HCC). (nih.gov)

The antibody was purified using epitope specific phosphopeptide. (abcam.com)
The antibody against non phosphopeptide was removed by chromatography using non phosphopeptide corresponding to the phosphorylation site. (abcam.com)
The antibody was affinity purified from rabbit antiserum by affinity chromatography using epitope-specific phosphopeptide. (abcam.com)
Each kit contains a total-ErbB-2 antibody that recognizes ErbB-2 regardless of its phosphorylation state. (activemotif.com)
The total-ErbB-2 antibody supplied in the FACE ErbB-2 Kits recognizes ErbB-2 protein regardless of its phosphorylation site. (activemotif.com)
Raptor was absent in this schema, so when TBK1 was examined against immunoprecipitated mTOR complexes, phosphorylation was measured with an antibody particular for phospho-Ser2159 mTOR. (healthweblognews.info)

Altered phosphorylation of CREB phosphopeptide, a substrate of GSK3β, suggested that GSK3β may be the mediator of Cdk5 activity in kinesin-mediated axonal transport. (frontiersin.org)
d) Alignment of the principal amino sequence of the phosphorylation sites determined by mass spectrometry with the most well-liked TBK1 substrate consensus sequence. (healthweblognews.info)
Three of the six phosphorylation sites had either leucine or isoleucine at the +1 position relative to the phosphorylation site, which matches the preferred substrate motif for TBK119,20 (Fig.?1d). (healthweblognews.info)

Custom Workflow for the Confident Identification of Sulfotyrosine-Containing Peptides and Their Discrimination from Phosphopeptides. (liverpool.ac.uk)
Glycophorin A was the most affected protein in sickle RBCs by this ERK1/2 pathway, which contained 12 unique phosphorylated peptides, suggesting that in addition to its effect on sickle RBC adhesion, increased glycophorin A phosphorylation via the ERK1/2 pathway may also affect glycophorin A interactions with band 3, which could result in decreases in both anion transport by band 3 and band 3 trafficking. (duke.edu)
phosphorylation of mutant 6His-M-1-I-116 peptides shows that CPK16 is able to phosphorylate the ACA8 N-terminus at Ser19 and at Ser22. (unimi.it)
Peptides with phosphorylation to serines, threonines or tyrosines are especially used in phosphoproteomics studies. (panatecs.com)

This is accomplished through phosphopeptide mapping, which involves digestion of a radioactively labeled protein, separation of phosphopeptide products, and finally analysis via high-performance liquid chromatography (HPLC) or mass spectrometry. (wikipedia.org)

Identification of in vitro 40 S ribosomal S6 phosphorylation sites. (concordia.ca)

Analysis of phosphopeptides can provide information about which amino acids are phosphorylated and how many sites on the primary sequence are phosphorylated. (wikipedia.org)
Mass spectrometry analysis of phosphopeptides was used to determine two MPK phosphorylation sites in ZAT10. (springer.com)
Multiple phosphorylation sites are identified in human TTP, but it is difficult to assign major vs. minor phosphorylation sites. (usda.gov)
Results: We identified two novel sites of serine phosphorylation at S63 and S262. (researchsquare.com)
Scoring and identifying organism-specific functional patterns and putative phosphorylation sites in protein sequences using mutual information. (mpg.de)
We have utilized mass spectrometry of purified bovine papillomavirus E1 protein to identify and characterize new sites of phosphorylation. (biomedcentral.com)
Five new and two previously known phosphorylation sites were identified. (biomedcentral.com)
A number of individual phosphorylation sites on BPV E1 have been identified by several groups, but no systematic effort to identify all of the phosphorylated amino acid positions of this protein has been undertaken. (biomedcentral.com)
Here we report five previously unidentified phosphorylation sites and confirm two known sites, identified by a combination of mass spectrometry (MS) methods. (biomedcentral.com)
Open in a separate window Figure 1 IRS-1 phosphopeptides containing multiple monophosphorylation sites. (iassist2012.org)
In comparison, there are a larger number of phosphorylation sites with significant changes in patient samples. (genomeweb.com)
These results indicate that regulation of synthase activity in the hepatocyte involves changes in phosphorylation at multiple sites. (umn.edu)
The level of protein phosphorylation at specific sites varies from less than 1% to greater than 90%, depending on conditions [ 26 ]. (biomedcentral.com)
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)
c) Domain framework of Raptor displaying the positions of the phosphorylation sites determined by mass spectrometry. (healthweblognews.info)
The phosphorylation sites corresponded to Ser44, Ser122, Ser836, Ser877 and Ser982 (Table?1 and Fig.?1c). (healthweblognews.info)
It may therefore be that the TBK1-dependent phosphorylation sites that match the motif are regulated by increases in TBK1 activity, whereas the others may be regulated BML-275 tyrosianse inhibitor by changes in TBK1 binding to Raptor. (healthweblognews.info)
Table 1 Phosphorylation Sites Identified Using Mass Spectrometry. (healthweblognews.info)

These analyses suggest that the tryptic peptide containing S90 and S93 is a major phosphopeptide in human TTP. (usda.gov)
Peptide sequence around phosphorylation site of threonine 200 (M-K-T(p)-V-C) derived from human CaMKIV. (genetex.com)
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)

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)
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)

Strong anion exchange-mediated phosphoproteomics reveals extensive human non-canonical phosphorylation. (liverpool.ac.uk)
A multipathway phosphopeptide standard for rapid phosphoproteomics assay development. (stanford.edu)
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)

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)

To mimic phosphorylation of the ACA8 N-terminus, each of the serines found to be phosphorylated in those studies (Ser19, Ser22, Ser27, Ser29, Ser57, and Ser99) has been mutated to aspartate. (unimi.it)

pp60c-src variants containing lesions that affect phosphorylation at tyrosines 416 and 527. (wikidata.org)

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)
For most of these approaches, three different quantitative methods (SILAC, iTRAQ, and label-free quantitation) can be introduced at steps indicated by "Q" to determine changes of phosphorylation between two or more samples. (yale.edu)

This analysis yielded 7868 quantifiable phosphopeptides and numerous up- or down-regulated candidates. (nih.gov)
Analysis using site-specific anti-phosphopeptide antibodies. (concordia.ca)
Recent advances in methodology have made phosphopeptide analysis a tractable problem for many proteomics researchers. (stanford.edu)
Proteomic analysis of ERK1/2-mediated human sickle red blood cell membrane protein phosphorylation. (duke.edu)

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 study generated additional information on TTP phosphorylation using phosphopeptide mapping and mass spectrometry. (usda.gov)
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)

In vivo phosphorylation of liver glycogen synthase. (umn.edu)
Incubation of in vivo phosphorylated synthase with endogenous synthase phosphatase to convert synthase D to synthase R resulted in removal of phosphate from all of the labeled phosphopeptides. (umn.edu)
Dive into the research topics of 'In vivo phosphorylation of liver glycogen synthase. (umn.edu)

The trypsin-derived [32P]phosphopeptides were separated by high pressure liquid chromatography (HPLC). (umn.edu)
When the HPLC [32P]phosphopeptide profile of synthase from glucagon-treated animals was compared with that of controls, there were only minor differences in the two profiles. (umn.edu)

We have used this cell line to investigate whether this temperature-dependent switching of function involves changes in the phosphorylation of p53 protein. (dundee.ac.uk)

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)

Protein phosphorylation is a very important and frequent post-translational modification that can impact a protein's localization, stability, and whether or not it can dimerize or form stable bonds with other substances. (wikipedia.org)

LC-MS/MS analyses identified 2755 and 3191 phosphopeptides from experiments with activator or inhibitor of PKA. (nih.gov)
Background: The phosphorylation of NF-kappaB inhibitor alpha (IκBα) protein is pivotal to the regulation of NF-κB transcription factor activity in the cell. (researchsquare.com)

The E1 helicase is regulated at least in part by protein phosphorylation, however no systematic approach to phosphate site mapping has been attempted. (biomedcentral.com)
A phosphate site map was created and used to develop a general model for the role of phosphorylation in E1 function. (biomedcentral.com)
A BPV E1 phosphate map and a discussion of the possible role of phosphorylation in E1 function are presented. (biomedcentral.com)

MS also identified a major phosphopeptide associated with the first zinc-finger region. (usda.gov)

Mutant TTP with an S197 mutation resulted in another major phosphopeptide peak being eluted earlier than the wild-type. (usda.gov)

phosphorylation of Tyr216 can enhance enzyme activity, and conversely phosphorylation of Ser9 is inhibitory. (frontiersin.org)
The possible physiological implications of the subtle modulation of ACA8 activity by phosphorylation of its N-terminus are discussed. (unimi.it)
The phosphopeptides enriched in the third reaction were presumed to be due to TBK1 activity. (healthweblognews.info)

Inhibition of Cdk5 by olomoucine enhanced phosphorylation of kinesin, suggesting that the effect was mediated by negative regulation of other kinases. (frontiersin.org)

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

Mutant TTP with S90 and S93 mutations resulted in the disappearance of a major phosphopeptide peak. (usda.gov)
At 32 degrees C and 37 degrees C phosphopeptide maps of the 'wild-type' and mutant protein were identical. (dundee.ac.uk)

MPKs can directly modulate gene expression by the phosphorylation of transcription factors. (springer.com)

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

We investigated the new phosphorylation site for IκBα and identified its biological function in breast cancer cells. (researchsquare.com)

Additional mutations at S186, S296 and T271 exhibited little effect on phosphopeptide profiles. (usda.gov)

11. Antion MD, Hou L, Wong H, Hoeffer CA, Klann E (2008) mGluR-dependent long-term depression is associated with increased phosphorylation of S6 and synthesis of elongation factor 1A but remains expressed in S6K-deficient mice. (concordia.ca)

Increased phosphorylation of LMNA and NIPA was confirmed using specific antibodies. (nih.gov)
Non-phospho specific antibodies were removed by chromatogramphy using non-phosphopeptide. (genetex.com)

C ) Phosphorylation of Pex14 upon treatment with oxidative agents. (elifesciences.org)
D ) Left, Time course of Pex14 phosphorylation upon H 2 O 2 treatment. (elifesciences.org)
There also was only a modest change in the [32P]phosphopeptide profile with glucose treatment when compared with that of controls. (umn.edu)

Fast Activated Cell-based ELISA (FACE™) Kits provide a simple, sensitive method for detecting protein phosphorylation directly in the cell, without making extracts or performing electrophoresis and membrane blotting. (activemotif.com)

Anatomy18

Organisms10

Chemicals and Drugs155

Analytical, Diagnostic and Therapeutic Techniques and Equipment26

Phenomena and Processes44

Disciplines and Occupations1

Technology, Industry, Agriculture1

Information Science3

Proteins13

Kinase10

Residues5

Synthetic phosphopeptide1

Aberrant phosphorylation1

Antibody6

Substrate3

Peptides4

High-performance li1

Identification1

Sites18

Peptide3

Assays2

Phosphoproteomics3

Enrichment1

Serines1

Tyrosines1

Quantitative2

Analysis4

Mass3

Vivo3

HPLC2

Involves1

Assay1

Substances1

Inhibitor2

Phosphate3

Zinc1

Mutation1

Activity3

Regulation1

Tissue2

Mutant2

Transcription1

Discovery2

Biological1

Effect1

Associated with increased1

Specific2

Treatment3

Cell1