Quantification of change in phosphorylation of BCR-ABL kinase and its substrates in response to Imatinib treatment in human chronic myelogenous leukemia cells

Liang, Xiquan; Hajivandi, Mahbod; Veach, Darren R.; Wisniewski, David; Clarkson, Bayard D.; Resh, Marilyn D.; Pope, R. Marshall

doi:10.1002/pmic.200600109

Cited by 44 publications

(30 citation statements)

References 36 publications

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“…8 Diminished SHIP1 activity or expression has been observed in human inflammatory diseases 9 and hematopoietic malignancies. [10][11][12][13] Because dysregulated activation of the PI3K pathway contributes to inflammatory/immune disorders and cancer, intense efforts have been invested into the development of inhibitors of PI3K itself, as well as downstream protein kinases. [14][15][16][17] The precedent for discovery and biologic efficacy of kinase inhibitors is well established, and a number of promising new PI3K isoform-specific inhibitors have recently been developed and used in mouse models of inflammatory disease 18,19 and glioma 20 with minimal toxicities.…”

Section: Introductionmentioning

confidence: 99%

Small-molecule agonists of SHIP1 inhibit the phosphoinositide 3-kinase pathway in hematopoietic cells

Ong

Ming-Lum

Nodwell³

et al. 2007

Blood

142

178

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Because phosphoinositide 3-kinase (PI3K) plays a central role in cellular activation, proliferation, and survival, pharmacologic inhibitors targeting components of the PI3K pathway are actively being developed as therapeutics for the treatment of inflammatory disorders and cancer. These targeted drugs inhibit the activity of either PI3K itself or downstream protein kinases. However, a previously unexplored, alternate strategy is to activate the negative regulatory phosphatases in this pathway. The SH2-containing inositol-5-phosphatase SHIP1 is a normal physiologic counterregulator of PI3K in immune/hematopoietic cells that hydrolyzes the PI3K product phosphatidylinositiol-3,4,5-trisphosphate (PIP 3 ). We now describe the identification and characterization of potent and specific small-molecule activators of SHIP1. These compounds represent the first small-molecule activators of a phosphatase, and are able to activate recombinant SHIP1 enzyme in vitro and stimulate SHIP1 activity in intact macrophage and mast cells. Mechanism of activation studies with these compounds suggest that they bind a previously undescribed, allosteric activation domain within SHIP1. Furthermore, in vivo administration of these compounds was protective in mouse models of endotoxemia and acute cutaneous anaphylaxis, suggesting that SHIP1 agonists could be used therapeutically to inhibit the PI3K pathway. IntroductionIn response to extracellular signals, phosphoinositide 3-kinase (PI3K) becomes activated and phosphorylates phosphatidylinositol-4,5-bisphosphate (PI-4,5-P 2 ) within the plasma membrane to generate phosphatidylinositol-3,4,5-bisphosphate (PIP 3 ). PIP 3 then initiates a cascade of downstream signaling pathways by interacting with pleckstrin homology (PH) domain-containing proteins, such as protein kinase B (PKB, also known as Akt), that regulate cellular activation, proliferation and/or survival, depending on the cell type and stimulus. 1 Cellular levels of PIP 3 are normally tightly regulated by both PI3K and the 5Ј inositol phosphatases SHIP1 (SH2 domain-containing inositol phosphatase) and SHIP2, as well as the 3Ј inositol phosphatase PTEN, which dephosphorylates PIP 3 . 2,3 Of these, SHIP1 is unique in that its expression is restricted primarily to immune and hematopoietic cells. 2,4 SHIP's role in immune cell homeostasis is shown both by the myeloproliferative syndrome observed in SHIP1 Ϫ/Ϫ mice, as well as the hypersensitivity of SHIP1 Ϫ/Ϫ mice and cells to immune stimulation. 5,6 SHIP1 mediates signaling from the inhibitory Fc␥RIIB receptor, 7 and is important in terminating signal transduction from activating immune/hematopoietic cell receptor systems. 8 Diminished SHIP1 activity or expression has been observed in human inflammatory diseases 9 and hematopoietic malignancies. [10][11][12][13] Because dysregulated activation of the PI3K pathway contributes to inflammatory/immune disorders and cancer, intense efforts have been invested into the development of inhibitors of PI3K itself, as well as downstream protein kinas...

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Section: Introductionmentioning

confidence: 99%

Small-molecule agonists of SHIP1 inhibit the phosphoinositide 3-kinase pathway in hematopoietic cells

Ong

Ming-Lum

Nodwell³

et al. 2007

Blood

142

178

View full text Add to dashboard Cite

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“…There is increasing evidence in the literature that SILAC is an excellent approach to study differential protein expression and posttranslational modification, especially phosphorylation [29,[33][34][35][36][37]. In this study, we combined SILAC with MMC and MMO to quantify the dynamic changes of tyrosine phosphorylation in response to EGF stimulation in HeLa cells.…”

Section: Isolation Of Phosphopeptide(s) From a Biological Sample Usinmentioning

confidence: 99%

“…Approximately one-third of the peptide extract was used for protein identification using nanoelectrospray LC-MS (Q-TOF premier; Waters Corporation, Milford, MA) or MALDI-TOF-TOF (4700 Proteomics Analyzer; Applied Biosystems) [29]. The rest of the sample was incubated with 250 g of MMC-Fe 3ϩ or MMO (TiO 2 ) for 10 min in 24 L of binding buffer to isolate phosphopeptides.…”

Section: Silac Labeling and Isolation Of Phosphopeptides For Lc-ms Anmentioning

confidence: 99%

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Quantitative comparison of IMAC and TiO₂ surfaces used in the study of regulated, dynamic protein phosphorylation

Liang¹,

Fonnum²,

Hajivandi³

et al. 2007

J. Am. Soc. Mass Spectrom.

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Protein phosphorylation regulates many aspects of cellular function, including cell proliferation, migration, and signal transduction. An efficient strategy to isolate phosphopeptides from a pool of unphosphorylated peptides is essential to global characterization using mass spectrometry. We describe an approach employing isotope tagging reagents for relative and absolute quantification (iTRAQ) labeling to compare quantitatively commercial and prototypal immobilized metal affinity chelate (IMAC) and metal oxide resins. Results indicate a prototype iron chelate resin coupled to magnetic beads outperforms either the Ga 3ϩ -coupled analog, Fe 3ϩ , or Ga 3ϩ -loaded, iminodiacetic acid (IDA)-coated magnetic particles, Ga 3ϩ -loaded Captivate beads, Fe 3ϩ -loaded Poros 20MC, or zirconium-coated ProteoExtract magnetic beads. For example, compared with Poros 20MC, the magnetic metal chelate (MMC) studied here improved phosphopeptide recovery by 20% and exhibited 60% less contamination from unphosphorylated peptides. With respect to efficiency and contamination, MMC performed as well as prototypal magnetic metal oxide-coated (TiO 2 ) beads (MMO) or TiO 2 chromatographic spheres, even if the latter were used with 2,5-dihydroxybenzoic acid (DHB) procedures. Thus far, the sensitivity of the new prototypes reaches 50 fmol, which is comparable to TiO 2 spheres. In an exploration of natural proteomes, tryptic (phospho)peptides captured from stable isotopic labeling with amino acids in cell culture (SILAC)-labeled immunocomplexes following EGF-treatment of 5 ϫ 10 7 HeLa cells were sufficient to quantify stimulated response of over 60 proteins and identify 20 specific phosphorylation sites. (J Am Soc Mass Spectrom 2007, 18, 1932-1944) © 2007 American Society for Mass Spectrometry C haracterization of protein phosphorylation status, especially following stimulated transduction events, can provide mechanistic insights into the biological basis of signaling, cell cycle progression, adhesion, migration, and numerous other functions. Nowadays, the identification of phosphorylation sites in a complex milieu is carried out mainly by mass spectrometry. However, the sensitivity of analysis is largely hindered by low stoichiometry of phosphorylation, the reversible nature of the modification, and relatively weak ionization of phosphopeptides. It has been noted that enrichment of phosphopeptides from a pool of unphosphorylated peptides dramatically improves the success frequency for characterization.Several methods for enriching phosphorylated peptides have been reported, including chemical derivatization of phospho-residues [1-5], antibody-based capture, immobilized metal affinity chromatography (IMAC) [6 -10], enrichment on metal oxide surfaces [11][12][13], strong cation exchange chromatography [14,15], and phosphoramidate chemistry [16,17]. For example, anti-phosphotyrosine antibodies have been used successfully to enrich tyrosine phosphorylated proteins and peptides [18 -22]. Some authors report hundreds of phosphopeptides are identi...

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“…Mass spectrometry (MS)-based quantitative phosphoproteomics has been a valuable tool to decipher signaling pathways initiated by a given tyrosine kinase (6). Among these, the stable isotope labeling with amino acids in cell culture (SILAC) has been used for the identification of oncogenic tyrosine kinase signaling such as HER2 (7) and Bcr-Abl (8). Here, we used this approach to address Src oncogenic signaling in advanced CRC cells.…”

Section: Introductionmentioning

confidence: 99%

Quantitative Phosphoproteomics Reveals a Cluster of Tyrosine Kinases That Mediates Src Invasive Activity in Advanced Colon Carcinoma Cells

et al. 2009

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The nonreceptor tyrosine kinase Src is frequently overexpressed and/or activated in human colorectal carcinoma (CRC), and its increased activity has been associated with a poor clinical outcome. Src has been implicated in growth and invasion of these cancer cells by still not well-known mechanisms. Here, we addressed Src oncogenic signaling using quantitative phosphoproteomics. Src overexpression increased growth and invasiveness of metastatic SW620 CRC cells. Stable isotope labeling with amino acids in cell culture in combination with liquid chromatography tandem mass spectrometry allowed the identification of 136 proteins which exhibited a significant increase in and/or association with tyrosine phosphorylation upon Src expression. These mainly include signaling, cytoskeleton, and vesicular-associated proteins. Interestingly, Src also phosphorylated a cluster of tyrosine kinases, i.e., the receptors Met and EphA2, the cytoplasmic tyrosine kinase Fak, and pseudo-tyrosine kinase SgK223, which were required for its invasive activity. Similar results were obtained with metastatic Colo205 CRC cells that exhibit high endogenous Src activity. We concluded that Src uses a tyrosine kinases network to promote its invasive activity in CRC and this implicates a reverse signaling via tyrosine kinase receptors. Targeting these tyrosine kinases may be of significant therapeutic value in this cancer. [Cancer Res 2009;69(6):2279-86]

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Quantification of change in phosphorylation of BCR-ABL kinase and its substrates in response to Imatinib treatment in human chronic myelogenous leukemia cells

Cited by 44 publications

References 36 publications

Small-molecule agonists of SHIP1 inhibit the phosphoinositide 3-kinase pathway in hematopoietic cells

Small-molecule agonists of SHIP1 inhibit the phosphoinositide 3-kinase pathway in hematopoietic cells

Quantitative comparison of IMAC and TiO₂ surfaces used in the study of regulated, dynamic protein phosphorylation

Quantitative Phosphoproteomics Reveals a Cluster of Tyrosine Kinases That Mediates Src Invasive Activity in Advanced Colon Carcinoma Cells

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Quantification of change in phosphorylation of BCR-ABL kinase and its substrates in response to Imatinib treatment in human chronic myelogenous leukemia cells

Cited by 44 publications

References 36 publications

Small-molecule agonists of SHIP1 inhibit the phosphoinositide 3-kinase pathway in hematopoietic cells

Small-molecule agonists of SHIP1 inhibit the phosphoinositide 3-kinase pathway in hematopoietic cells

Quantitative comparison of IMAC and TiO2 surfaces used in the study of regulated, dynamic protein phosphorylation

Quantitative Phosphoproteomics Reveals a Cluster of Tyrosine Kinases That Mediates Src Invasive Activity in Advanced Colon Carcinoma Cells

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Quantitative comparison of IMAC and TiO₂ surfaces used in the study of regulated, dynamic protein phosphorylation