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Antibodies, Phospho-Specific (0):
Recent article
Antibodies, Phospho-Specific
Biology Direct (6)1 2011
Abstract
We examined the specificity of several commercially available anti-phosphorylated GSK-3 antibodies. We show that antibodies raised to peptides containing the phosphorylated Ser21/9 epitope crossreact with unidentified antigens that are highly expressed by mitotic cells and that mainly localise to sp...
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PMID: 21261990
PDF is available here.
PDF is available here.
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Biochemical Journal (431)2 2010
Abstract
We studied TBC1D1 phosphorylation on three predicted AMPK (AMP-activated protein kinase) phosphorylation sites (Ser231, Ser660 and Ser700) and one predicted Akt phosphorylation site (Thr590) in control mice, AMPKα2 inactive transgenic mice (AMPKα2i TG) and Akt2-knockout mice (Akt2 KO). Muscle cont...
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PMID: 20701589
PDF is available here.
PDF is available here.
Amino Acids (38)4 2010
Abstract
Recent studies showed that a combination of carbohydrate and protein was more effective than carbohydrate alone for replenishing muscle glycogen after exercise. However, it remains to be unclear whether the source or degree of hydrolysis of dietary protein influences post-exercise glycogen accumulat...
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PMID: 19593593
PDF is available here.
PDF is available here.
Journal of Biochemistry (147)4 2010
Abstract
We have identified protein phosphatase 1alpha (PP1alpha) as a novel PTP-PEST binding protein, and then we aimed to determine how PP1alpha contributes to the phosphorylation at Ser39 of PTP-PEST, whose phosphorylation suppresses PTP-PEST enzymatic activity. The HEK 293 cells overexpressing exogenous...
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PMID: 19919952
PDF is available here.
PDF is available here.
Molecular and Cellular Biochemistry (336)1-2 2010
Abstract
Protein tyrosine phosphorylation is a key event accompanying sperm capacitation. Although this signaling cascade generates an array of tyrosine-phosphorylated polypeptides, their molecular characterization is still limited. It is necessary to differentiate the localization of the tyrosine-phosphoryl...
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PMID: 19802524
PDF is available here.
PDF is available here.
Abstract
We identified the activation of the mammalian target of rapamycin (mTORC)1 pathway in 29 cases of AIDS-related lymphoma. These cases represented a diverse spectrum of histological types of non-Hodgkin lymphoma (24 cases) and classic Hodgkin lymphoma (five cases). mTORC1 was also activated in the hyp...
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PMID: 19608873
PDF is available here.
PDF is available here.
Journal of Biological Chemistry (284)31 2009
Abstract
We applied a novel approach to select monoclonal phosphospecific antibodies directly from the full-length protein and without up-front phosphoamino acid identification. We chose as antigen GRASP65, a well studied Golgi phosphoprotein. Bacterially produced full-length protein was first incubated with...
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PMID: 19473967
PDF is available here.
PDF is available here.
Methods in Molecular Biology (527)527 2009
Abstract
In view of the importance of information transfer mediated throughout the cell by recognition, phosphorylation or dephosphorylation of kinases, their adapters, or substrates, this method was developed. The method provides a potent research tool for rapidly generating and testing these substrates as...
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PMID: 19241019
PDF is available here.
PDF is available here.
Methods in Molecular Biology (527)527 2009
Abstract
We have described in detail two methods for differential analysis of protein tyrosine phosphorylation in cells by (1) using a single fluorescent protein capture format on membrane array and (2) a competitive protein capture method on glass surface array....
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PMID: 19241018
PDF is available here.
PDF is available here.
Abstract
We have recently demonstrated that BRCA1 reduces acetyl-CoA-carboxylase alpha (ACCA) activity through its phospho-dependent binding to ACCA, and further established that the phosphorylation of the Ser1263 of ACCA is required for this interaction. Here, to gain more insight into the cellular conditio...
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PMID: 19061860
PDF is available here.
PDF is available here.
Folia Biologica (Prague) (55)4 2009
Abstract
We have outlined detailed procedures for fibronectin-replating experiment, biochemical examination of the phosphotyrosine content of cellular proteins by immunoblotting using phosphorylation-specific antibodies or immunoprecipitation and analysis with general phosphotyrosine antibodies. Despite grea...
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PMID: 19691923
PDF is available here.
PDF is available here.
Abstract
Protein phosphorylation is a versatile posttranslational modification that can regulate nuclear receptor function. Although the precise role of receptor phosphorylation is not fully understood, it appears that it functions to direct or refine receptor activity in response to particular physiological...
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PMID: 19117148
PDF is available here.
PDF is available here.
Abstract
We present here four methods to isolate the TCR in a native form and details to analyse it by BN-PAGE....
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PMID: 19048218
PDF is available here.
PDF is available here.
Abstract
We have to use immunohistochemistry. The protocol described here allows the detection of phosphorylated histones in tissue-cultured cells and tissue sections by fluorescent or bright-field immunostaining analysis. Here we used a serine 10 specific P-histone H3 antibody to determine the localization...
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PMID: 19381929
PDF is available here.
PDF is available here.
Abstract
We measured phosphorylation with at least two different antibodies per site and demonstrate that for S2808 results were highly antibody-dependent and two out of three S2808 antibodies did not accurately report phosphorylation level. The RyR was substantially phosphorylated in quiescent rat cardiomyo...
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PMID: 18755143
PDF is available here.
PDF is available here.
Abstract
We compared data generated by WB or Phospho-Flow regarding the kinetics and degree of phosphorylation of membrane proximal TCR signaling molecules. Phosphorylation events in Jurkat T cells were triggered by anti-CD3 stimulation (OKT3) or by oxidative stress (H(2)O(2)) and were analyzed by Phospho-Fl...
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PMID: 18548611
PDF is available here.
PDF is available here.
Molecular and Cellular Biochemistry (316)1-2 2008
Abstract
We have recently produced an anti-[pSer13]-Cdc37 antibody which specifically recognizes Cdc37 that is phosphorylated on Ser 13 [Miyata Y. & Nishida E. FEBS J. 274:5690-5703, 2007]. Here we investigated CK2 activity both in vitro and in cultured cells by using anti-[pSer13]-Cdc37 antibody. Immunoblot...
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PMID: 18566753
PDF is available here.
PDF is available here.
Molecular Endocrinology (22)8 2008
Abstract
We demonstrated that GR transcriptional activation is greatest when the relative phosphorylation of S211 exceeds that of S226. Consistent with this finding, a replacement of S226 with an alanine enhances GR transcriptional response. Using a battery of compounds that perturb different signaling pathw...
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PMID: 18483179
PDF is available here.
PDF is available here.
Journal of Cell Science (121)Pt 14 2008
Abstract
We sought to identify a PE-induced phosphorylation site in AFAP-110 and determine whether phosphorylation is linked to the formation of podosomes. Mutational analysis revealed Ser277 of AFAP-110 to be phosphorylated in PE-treated cells. The use of a newly generated, phospho-specific antibody directe...
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PMID: 18577577
PDF is available here.
PDF is available here.
Abstract
We raised antibodies directed against different mitotic phosphorylation sites of human Cdc25C, and characterized the phosphorylated species detectable in HeLa cells. In the present study, we provide first-time evidence for the existence of multiple species of Cdc25C in mitotic cell extracts, includi...
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PMID: 18384749
PDF is available here.
PDF is available here.
Nucleic Acids Research (36)3 2008
Abstract
We found that phosphorylation of PAP was increased upon growth stimulation and that the mitogen-activated protein kinase ERK was responsible for the increase in phosphorylation. We identified serine 537 of PAP as a unique phosphorylation site by ERK. PAP phosphorylation of serine 537 by ERK increase...
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PMID: 18084034
PDF is available here.
PDF is available here.
Nucleic Acids Research (36)2 2008
Abstract
We have previously shown that human exonuclease 1 (hEXO1) is phosphorylated in response to agents stalling DNA replication and that hEXO1 consequently undergoes ubiquitination and degradation in a proteasome-dependent manner. In the present study, we have addressed the identity of the pathway transd...
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PMID: 18048416
PDF is available here.
PDF is available here.
Abstract
We investigated myocytes from CaMKIIdelta(C) transgenic (TG) mice (cytosolic localized CaMKII) having heart failure vs. wild-type littermates (WT). Furthermore, mouse and rabbit myocytes overexpressing CaMKIIdelta(C) using adenovirus-mediated gene transfer (vs. LacZ control) were investigated. Fract...
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PMID: 17950750
PDF is available here.
PDF is available here.
Proteomics (7)24 2007
Abstract
We studied the effects of a p38 MAP kinase inhibitor, PD169316, on cardioprotection, protein expression, and tyrosine phosphorylation, in a rabbit model of 1 h of (MI) and 3 h of (R). PD169316 administered just before (R) significantly reduced myocardial neutrophil accumulation, necrosis area (28.4...
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PMID: 18072194
PDF is available here.
PDF is available here.
Abstract
We utilize a peptide library and bioinformatic approach to predict likely substrates of IKKbeta. This approach predicted Ser381 of the K63 deubiquitinase A20 as a likely site of IKKbeta phosphorylation. While A20 is a known negative regulator of innate immune signaling pathways, the mechanisms regul...
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PMID: 17709380
PDF is available here.
PDF is available here.
Growth Factors (25)5 2007
Abstract
We have previously shown that phosphorylation of tyrosine1056 of ErbB4 is critical for it to inhibit colony formation on plastic by human tumor cell lines. Thus, an antibody directed against this site would be useful for studying ErbB4 signaling and coupling to biological responses. Here, we demonst...
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PMID: 18236211
PDF is available here.
PDF is available here.
Abstract
Histone Deacetylase 1 (HDAC1) is a transcriptional regulator associated with proliferation, apoptosis, and tumorigenesis, although its precise cellular role is unclear. HDAC1 was previously characterized as a phosphoprotein where mutation of phosphorylated S421 and S423 resulted in a loss of deacety...
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PMID: 17643391
PDF is available here.
PDF is available here.
Cell Calcium (42)3 2007
Abstract
We report here that IP3R2 is only very weakly phosphorylated in response to PKA activation and is probably not a physiological substrate for this kinase. IP3R3, however, is known to be phosphorylated by PKA at three sites (S916, S934, and S1832) and, thus, we examined how phosphorylation of these si...
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PMID: 17257671
PDF is available here.
PDF is available here.
Cellular Signalling (19)9 2007
Abstract
We demonstrate that GFP-PKCepsilon concentrates at a perinuclear site in living fibroblasts and that cell passage induces rapid translocation of PKCepsilon to the periphery where it appears to colocalise with F-actin. When newly passaged cells have adhered and are proliferating again, GFP-PKCepsilon...
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PMID: 17611075
PDF is available here.
PDF is available here.
Abstract
This unit describes three procedures for measurement of interleukin-21 (IL-21). The first employs the use of an antibody sandwich ELISA. An alternative procedure measures proliferative responses of T cells to a combination of IL-21 and IL-15 using CFSE. Finally, a method to assess IL-21-induced tyro...
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PMID: 18432995
PDF is available here.
PDF is available here.
Abstract
This protocol describes methods for monitoring intracellular phosphorylation-dependent signaling events on a single-cell basis. This approach measures cell signaling by treating cells with exogenous stimuli, fixing cells with formaldehyde, permeabilizing with methanol, and then staining with phospho...
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PMID: 18432997
PDF is available here.
PDF is available here.
Biochemical Journal (405)1 2007
Abstract
We demonstrate that the metabolic sensing kinase AMPK (AMP-activated protein kinase) phosphorylates NKCC2 on Ser126 in vitro. Co-precipitation experiments indicated that there is a physical association between AMPK and the N-terminal cytoplasmic domain of NKCC2. Activation of AMPK in the MMDD1 (mous...
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PMID: 17341212
PDF is available here.
PDF is available here.
Journal of Biological Chemistry (282)24 2007
Abstract
We screened for potential ATM/ATR substrates using phospho-specific antibodies against known ATM/ATR substrates. We identified proteins cross-reacting to phospho-specific antibodies in response to DNA damage by mass spectrometry. We validated a subset of the candidate substrates to be phosphorylated...
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PMID: 17478428
PDF is available here.
PDF is available here.
Journal of Biological Chemistry (282)12 2007
Abstract
We used a polyreactive phosphospecific antibody (alpha-pDSQ) that recognizes a subset of phosphorylated Asp-Ser-Gln sequences to purify candidate ATM/ATR substrates. This led to the identification of phosphorylation sites in the carboxyl terminus of the minichromosome maintenance protein 3 (MCM3), a...
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PMID: 17244605
PDF is available here.
PDF is available here.
Abstract
We generated site-specific monoclonal antibodies that specifically react with phosphoserine 334. In differentiated U937 cells, which endogenously express C5aR, stimulation with low C5a concentrations resulted in a very rapid (t((1/2)) approximately 20 s), albeit transient, receptor phosphorylation....
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PMID: 17145764
PDF is available here.
PDF is available here.
FEBS Letters (581)2 2007
Abstract
We report here that PSF is phosphorylated in vivo to generate an epitope(s) that can be recognized by a monoclonal antibody specific for phosphorylated RS motifs within SR proteins. PSF can be phosphorylated by human and yeast SR kinases in vivo and in vitro at an isolated RS motif within its N term...
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PMID: 17188683
PDF is available here.
PDF is available here.
Abstract
Formation of the mammalian orofacial region involves multiple signaling pathways regulating sequential expression of and interaction between molecular signals during embryogenesis. The present study examined the expression patterns of members of the MAPK family in developing murine orofacial tissue....
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PMID: 17177285
PDF is available here.
PDF is available here.
Abstract
Protein phosphorylation plays important roles in various aspects of cellular events. Visualization of site-specific phosphorylation in cells is of great importance not only to analyze spatial and temporal distribution but also to investigate biological function. Now, site- and phosphorylation state-...
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PMID: 17948000
PDF is available here.
PDF is available here.
Abstract
This chapter will describe methods to assess the activities of protein kinases. Initial studies in the 1950s and 1960s in the field of glucose metabolism examined the activities of several highly specific protein and carbohydrate kinases in cell lysates or isolated cell fractions. As more protein ki...
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PMID: 18217691
PDF is available here.
PDF is available here.
Abstract
Synthetic peptides are widely used for production and analysis of antibodies as well as in the study of protein modification enzymes. To circumvent the technical challenges of the existing techniques regarding peptide quantization and normalization, a new method of producing peptide arrays has been...
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PMID: 17984527
PDF is available here.
PDF is available here.
Molecular and Cellular Biochemistry (295)1-2 2007
Abstract
We made a specific antibody against the phosphor-SAG(Thr10). Transient transfection experiment showed that SAG was phosphorylated at Thr10 which can be significantly inhibited by TBB, a relatively specific inhibitor of protein kinase CK2. To determine whether this SAG phosphorylation is stress-respo...
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PMID: 16874460
PDF is available here.
PDF is available here.
Development (133)21 2006
Abstract
We have examined the molecular underpinnings of IP3R1 function in eggs. Using mouse and Xenopus eggs, we show that IP3R1 is phosphorylated during both maturation and the first cell cycle at a MPM2-detectable epitope(s), which is known to be a target of kinases controlling the cell cycle. In vitro ph...
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PMID: 17038520
PDF is available here.
PDF is available here.
FEBS Letters (580)24 2006
Abstract
We have developed a generic assay to quantitatively measure phospho-specific interactions between a trafficking signal-phosphorylated SWTY peptide and 14-3-3 proteins or anti-phosphopeptide antibodies. Compared with a solution-based fluorescence anisotropy assay, our results support that the high th...
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PMID: 17011553
PDF is available here.
PDF is available here.
Abstract
We clarify this issue using an approach combining mass spectrometry and the use of specific antibodies against phosphorylated CDC25B residues. We report that MAPKAP kinase-2 phosphorylates CDC25B on multiple sites including S169, S323, S353 and S375, while p38SAPK phosphorylates CDC25B on S249. We s...
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PMID: 16861915
PDF is available here.
PDF is available here.
Abstract
Abstract
We find that all forms of polymerase II are present, but not enriched, within speckles. Inhibition of polymerase activity shows that speckles do not act as major storage sites for inactive polymerase II complexes but that they contain a stable pool of polymerase II phosphorylated on serine(2) residu...
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PMID: 16467386
PDF is available here.
PDF is available here.
Abstract
We show that Cdk1 phosphorylates beta-tubulin both in vitro and in vivo. Phosphorylation occurs on Ser172 of beta-tubulin, a site that is well conserved in evolution. Using a phosphopeptide antibody, we find that a fraction of the cell tubulin is phosphorylated during mitosis, and this tubulin phosp...
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PMID: 16371510
PDF is available here.
PDF is available here.
Abstract
We report here three novel epidermal growth factor-induced in vivo Raf-1 phosphorylation sites that mediate positive feedback Raf-1 regulation. Using mass spectrometry, we identified Raf-1 phosphorylation on three SP motif sites: S289/S296/S301 and confirmed their identity using two-dimensional-phos...
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PMID: 16407412
PDF is available here.
PDF is available here.
Proteomics (6)3 2006
Abstract
We transition from genomics to the challenges of the functional proteome, new tools to explore the expression of proteins within tissue are essential. We have developed a method of transferring proteins from a formalin fixed, paraffin embedded tissues section to a stack of membranes which is then pr...
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PMID: 16400680
PDF is available here.
PDF is available here.
FEBS Letters (580)2 2006
Abstract
We have previously identified Thr(199) as the major phosphorylation site of NPM mediated by CDK2/cyclin E (and A), and this phosphorylation is involved in the regulation of centrosome duplication. In this study, we further examined the effect of CDK2-mediated phosphorylation of NPM by using the anti...
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PMID: 16376875
PDF is available here.
PDF is available here.
EMBO Journal (25)1 2006
Abstract
We identified a key regulatory phosphorylation site, serine 519 and serine 526, in MAP3K MEKK2 and MEKK3, respectively. Mutation of this serine to an alanine severely impaired MEKK2/3 activation. We generated an anti-p-MEKK2/3 antibody and used this antibody to demonstrate that lipopolysaccharide in...
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PMID: 16362041
PDF is available here.
PDF is available here.
Abstract
The biological functions of mast cells are regulated by several protein kinases, including the tyrosine kinases Fyn, Lyn, Syk, and FAK and the serine/threonine kinases Akt and PKC alpha/beta. The mitogen-activated protein kinases extracellular signal-regulated kinases, JNK, and p38MAPK also play a s...
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PMID: 16110156
PDF is available here.
PDF is available here.
Abstract
Phospho-specific antibodies have become very useful reagents for study of signal transduction pathways. This chapter describes the production of phospho-specific antibodies and their use to assess individual phosphorylation events in vivo in cells. The first step involves the synthesis of peptides (...
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PMID: 16491617
PDF is available here.
PDF is available here.
Infection and Immunity (73)12 2005
Abstract
We showed that the target of the phosphatase activity is the ectodomain of CD36 at threonine-92 (Thr92). Mouse fibroblasts (NIH 3T3 cells) transfected with wild-type CD36 or a mutant protein in which Thr92 was substituted by Ala supported the rolling and adhesion of IRBCs. However, while the Src fam...
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PMID: 16299313
PDF is available here.
PDF is available here.
Aquatic Toxicology (75)3 2005
Abstract
We investigated the possible effects and mechanisms of action of TBBPA on the immune cells, the hemocytes, of the marine mussel Mytilus galloprovincialis. The results demonstrate that TBBPA in the low micromolar range induces hemocyte lysosomal membrane destabilization. The effect was reduced or pre...
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PMID: 16198432
PDF is available here.
PDF is available here.
Biochemical Journal (391)Pt 2 2005
Abstract
We have found that the Nogo-B spliceform becomes phosphorylated at Ser107 in response to lipopolysaccharide in RAW264 macrophages or anisomycin in HeLa cells. The phosphorylation is prevented by SB 203580, an inhibitor of SAPK2a (stress-activated protein kinase 2a)/p38a and SAPK2b/p38b, and does not...
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PMID: 16095439
PDF is available here.
PDF is available here.
Abstract
Nature Genetics (37)10 2005
Abstract
Abstract
We investigated the mechanisms involved in estradiol (E2)-induced SRC-3 phosphorylation and found that this occurs only when either activated estrogen receptor alpha (ERalpha) or activated ERbeta is present. Both the activation function 1 and the ligand binding domains of ERalpha are required for ma...
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PMID: 16135815
PDF is available here.
PDF is available here.
Molecular Cell (19)3 2005
Abstract
We demonstrate that the receptor for activated C kinase 1 (RACK1) serves as an adaptor for PKC-mediated JNK activation. Phosphorylation of JNK by PKC occurs on Ser129 and requires the presence of RACK1. Ser129 phosphorylation augments JNK phosphorylation by MKK4 and/or MKK7 and is required for JNK a...
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PMID: 16061178
PDF is available here.
PDF is available here.