PP58
目录号 : GC32835
PP58是吡啶[2,3-d]嘧啶化合物，抑制PDGFR，FGFR和Src家族活性的IC50值在纳摩尔级。
Cas No.:212391-58-7
Sample solution is provided at 25 µL, 10mM.
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Quality Control & SDS
- View current batch:
- Purity: >99.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Kinase experiment: | MEK1 and Aurora A activities are tested at 37 °C in a total volume of 30 μL. The kinases are assayed using 50 μM ATP and 1 μCi [γ-32P]ATP in the presence of different PP58 concentrations. Kinase substrate proteins included are 0.25 mg/mL inactive GST-ERK2 for MEK1 and 0.025 mg/mL kemptide for Aurora A, respectively. Reactions are stopped by addition of SDS sample buffer. Determination of IC50 [0–100%] values is performed using GraFit software[1]. |
References: [1]. Wissing J, et al. Chemical Proteomic Analysis Reveals Alternative Modes of Action for Pyrido[2,3-d]pyrimidine Kinase Inhibitors. Mol Cell Proteomics. 2004 Dec;3(12):1181-93. | |
PP58 is a pyrido[2,3-d]pyrimidine-based compound that inhibits PDGFR, FGFR and Src family activities with nanomolar IC50 values.
PP58 inhibits Src with a subnanomolar IC50 value in the assays. PP58 behaves as a titration reagent at higher Src protein concentrations. As analyzed by immunoblotting with specific antiserum, the PP58 matrix specifically depletes Src from total lysate, whereas binding to the PP58 beads is prevented when free inhibitor is included. The ectopically expressed FGFR1 receptor tyrosine kinase is specifically retained on PP58 beads. PP58 matrix could be a novel affinity reagent for the purification of cellular pyrido[2,3-d]pyrimidine inhibitor targets. PP58 affinity chromatography leads to the identification of protein kinases belonging to various different groups and families, indicating that the pyrido[2,3-d]pyrimidine inhibitor is not selective for a set of phylogenetically related members of the human kinome. The Ki values of PP58 for p38α and JNK2 are 3.8±1.9 nM and 0.32±0.04 μM, respectively. PP58 affinity matrix also serves as an efficient purification reagent for a variety of protein kinases, which lack this structural feature and have much lower affinities for the pyrido[2,3-d]pyrimidine inhibitor PP58. PP58 inhibits anisomycin activated p38 in a dose-dependent manner with an IC50 below 10 nM. LPS-stimulated TNF-α production is potently inhibited by PP58 with a cellular IC50 value of around 3 nM[1]. The T341M mutation abrogates the sensitivity to PP58 inhibition by increasing the cellular IC50 value of about 10 nM by more than 1000-fold. The cellular wild-type FGFR1 activity is potently inhibited by low nanomolar concentrations of PP58, whereas dramatic resistance formation is detected for the FGFR1-V561M mutant. PP58 inhibits CSK activity with an IC50 value of around 100 nM[2].
PP58 can exhibit some degree of selectivity at low nanomolar concentrations in vivo[1].
[1]. Wissing J, et al. Chemical Proteomic Analysis Reveals Alternative Modes of Action for Pyrido[2,3-d]pyrimidine Kinase Inhibitors. Mol Cell Proteomics. 2004 Dec;3(12):1181-93. [2]. Blencke S, et al. Characterization of a conserved structural determinant controlling protein kinase sensitivity to selective inhibitors. Chem Biol. 2004 May;11(5):691-701.
| Cas No. | 212391-58-7 | SDF | |
| Canonical SMILES | NCCOC1=CC=C(NC(N=C2N3C)=NC=C2C=C(C4=C(Cl)C=CC=C4Cl)C3=O)C=C1 | ||
| 分子式 | C22H19Cl2N5O2 | 分子量 | 456.32 |
| 溶解度 | Soluble in DMSO | 储存条件 | Store at -20°C |
| General tips | 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。 储备液的保存方式和期限：-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
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| Shipping Condition | 评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备RT，或根据请求配备蓝冰。 | ||
| 制备储备液 | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 2.1914 mL | 10.9572 mL | 21.9144 mL |
| 5 mM | 0.4383 mL | 2.1914 mL | 4.3829 mL |
| 10 mM | 0.2191 mL | 1.0957 mL | 2.1914 mL |
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| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
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| % DMSO % % Tween 80 % saline | ||||||||||
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计算结果:
工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。
1. 首先保证母液是澄清的；
2.
一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
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Characterization of two highly phosphorylated cytoskeleton-associated proteins, PP58 and pp60, in tumoricidal murine peritoneal macrophages and their comparison with vimentin
Mol Immunol 1988 Aug;25(8):785-94.PMID:3185571DOI:10.1016/0161-5890(88)90114-9.
Two TX-insoluble cytoskeleton-associated proteins, PP58 and pp60, become highly phosphorylated in tumoricidal murine peritoneal macrophages. Results suggest that PP58 (pI 5.00) is phosphovimentin because it is highly insoluble in TX, shares the same mol. wt as vimentin, has a more acidic isoelectric point than vimentin, is phosphorylated primarily at serine, and generates the same V-8 protease peptide map as vimentin. pp60 generates at slightly different peptide map than PP58 and has a slightly less acidic isoelectric point (pI 5.02) than PP58 (pI 5.00), but is similar to PP58 by being highly insoluble in TX and being phosphorylated primarily at serine residues. Pulse-chase experiments demonstrate that PP58 is not a precursor to or breakdown product of pp60, or vice versa because they show similar rates of [32P]-phosphate incorporation and turnover.
Phosphorylation of cytoskeleton-associated proteins, PP58 and pp60, in tumouricidal murine peritoneal macrophages
Immunol Cell Biol 1989 Oct;67 ( Pt 5):311-9.PMID:2613279DOI:10.1038/icb.1989.46.
Two highly phosphorylated vimentin-like proteins, PP58 and pp60, are expressed in macrophages activated in vivo to tumouricidal activity. Resident and elicited, non-tumouricidal peritoneal macrophages displayed low and intermediate levels of phosphorylated PP58 and pp60, respectively. C3H/HeN macrophages became tumouricidal after incubation with 0.1 micrograms/mL A23187 plus 10 nmol/L 12-phorbol 13-myristate acetate (PMA), or 0.1 micrograms/mL A23187 plus 100 ng/mL lipopolysaccharide (LPS), and displayed increased phosphorylation of PP58 and pp60. LPS non-responder C3H/HeJ macrophages were not tumouricidal nor did they show increased phosphorylation of PP58 and pp60 after incubation with LPS plus A23187 in vitro. C3H/HeJ macrophages, however, did become tumouricidal and expressed increased phosphorylation of PP58 and pp60 after incubation with A23187 and PMA. Addition of PGE2 (10(-8) mol/L), resulted in down-regulation of macrophage tumouricidal activity and decreased PP58 and pp60 phosphorylation, which was reversed by addition of indomethacin (10(-6) mol/L) to cultures with PGE2. Phosphorylation increased within 5 min after adding activating stimuli while incorporation of [35S]-methionine into a 58 kD protein did not occur until 6 h later. No 60 kD protein synthesis was detected during the first 8 h after adding activating stimuli, indicating that previously synthesized proteins were phosphorylated during macrophage activation. These results signify a physiological role for the phosphorylation of cytoskeleton-associated PP58 and pp60 during macrophage activation to tumour cytotoxicity.
Immunological characterization of the Epstein-Barr virus phosphoprotein PP58 and deoxyribonuclease expressed in the baculovirus expression system
Int J Cancer 1991 Jul 30;48(6):879-88.PMID:1650330DOI:10.1002/ijc.2910480615.
The open reading frames of the phosphoprotein PP58 (BMRFI) and the deoxyribonuclease (BGLF5) of the Epstein-Barr-virus (EBV) strain M-ABA were cloned in the baculovirus expression vectors pAc373 and pAc360 and expressed in the Spodoptera frugiperda (SF158) insect cells. The recombinant phosphoprotein PP58 expressed in SF158 cells was recognized by the anti-pp58 rabbit anti-sera which were generated by immunizing rabbits with a TrpE-BMRFI fusion protein expressed in E. coli. DNA-cellulose chromatography showed that the recombinant PP58 exhibited DNA-binding activities. Immunofluorescence, immunoblot and ELISA analysis indicated that sera from patients with nasopharyngeal carcinoma (NPC) contained antibodies against PP58. The recombinant EBV DNase expressed in SF158 cells was recognized by the anti-EBV DNase rabbit anti-sera which were generated by immunizing rabbits with a TrpE-C-terminal part of BGLF5 fusion protein expressed in E. coli. The anti-EBV DNase rabbit anti-sera recognized also a protein of about 52 kDa in the EBV-harboring human B-cell lines Raji, Jijoye, B95-8, M-ABA and BL74 induced by TPA and n-butyrate. The recombinant EBV DNase exhibited exonuclease and endonuclease activities, a requirement for magnesium, and a high pH optimum (8.0). Its enzyme activities could be inhibited by sera from NPC patients and anti-EBV DNase rabbit anti-sera. Comparable studies of Raji EBV-DNase and recombinant EBV-DNase implied that recombinant EBV-DNase could also be used in the enzyme activity assay for the detection of NPC. In contrast to the enzyme inhibition test, immunofluorescence and immunoblot analysis demonstrated that the recombinant EBV DNase exhibited only a weak immunological reaction with NPC sera.
Pillars article: the CD4 receptor is complexed in detergent lysates to a protein-tyrosine kinase (PP58) from human T lymphocytes. 1988
J Immunol 2010 Sep 1;185(5):2645-9.PMID:20724730doi
The CD4 (T4) antigen is a cell-surface glycoprotein that is expressed predominantly on the surface of helper T cells and has been implicated in the regulation of T-cell activation and in the associative recognition of class II antigens of the major histocompatibility complex. In addition, the CD4 antigen appears to serve as a receptor for the human immunodeficiency virus (HIV). An important question has been whether the CD4 receptor is linked to an intracellular mediator that could regulate the activation of the CD4+ subset. In this paper, we provide preliminary evidence that the CD4 receptor is complexed in detergent lysates to a protein-tyrosine kinase (PTK) of 55–60 kDa, which is expressed specifically in T cells. The PTK is the human analogue of the murine pp56LSTRA (pp56lck) and has significant homology with c-src, c-yes, and other members of the src family. The identification of the PTK associated with CD4 receptor was made by use of an antiserum to a synthetic peptide that was deduced from the DNA sequence of PTK. Two-dimensional nonequilibrium pH gradient gel electrophoresis/NaDodSO4/PAGE revealed the kinase to focus as a heterogeneous collection of spots in the pH range of 4.0–5.0. Furthermore, in vitro phosphorylation revealed the phosphorylation of two additional polypeptides at 40 and 80 kDa, in addition to the autophosphorylation of the PTK at 55–60 kDa. The potential importance of the association between the CD4 receptor and the PTK of T cells is discussed in relation to T-cell activation and HIV infectivity.
The CD4 receptor is complexed in detergent lysates to a protein-tyrosine kinase (PP58) from human T lymphocytes
Proc Natl Acad Sci U S A 1988 Jul;85(14):5190-4.PMID:2455897DOI:10.1073/pnas.85.14.5190.
The CD4 (T4) antigen is a cell-surface glycoprotein that is expressed predominantly on the surface of helper T cells and has been implicated in the regulation of T-cell activation and in the associative recognition of class II antigens of the major histocompatibility complex. In addition, the CD4 antigen appears to serve as a receptor for the human immunodeficiency virus (HIV). An important question has been whether the CD4 receptor is linked to an intracellular mediator that could regulate the activation of the CD4+ subset. In this paper, we provide preliminary evidence that the CD4 receptor is complexed in detergent lysates to a protein-tyrosine kinase (PTK) of 55-60 kDa, which is expressed specifically in T cells. The PTK is the human analogue of the murine pp56LSTRA (pp56lck) and has significant homology with c-src, c-yes, and other members of the src family. The identification of the PTK associated with CD4 receptor was made by use of an antiserum to a synthetic peptide that was deduced from the DNA sequence of PTK. Two-dimensional nonequilibrium pH gradient gel electrophoresis/NaDodSO4/PAGE revealed the kinase to focus as a heterogeneous collection of spots in the pH range of 4.0-5.0. Furthermore, in vitro phosphorylation revealed the phosphorylation of two additional polypeptides at 40 and 80 kDa, in addition to the autophosphorylation of the PTK at 55-60 kDa. The potential importance of the association between the CD4 receptor and the PTK of T cells is discussed in relation to T-cell activation and HIV infectivity.