DOCSLIB.ORG

Protein Tyrosine Phosphatase Receptor Type Γ Is a Functional Tumor Suppressor Gene Specifically Downregulated in Chronic Myeloid Leukemia

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Protein Tyrosine Phosphatase Receptor Type Γ Is a Functional Tumor Suppressor Gene Specifically Downregulated in Chronic Myeloid Leukemia Published OnlineFirst October 19, 2010; DOI: 10.1158/0008-5472.CAN-10-0258 Tumor and Stem Cell Biology Cancer Research Protein Tyrosine Phosphatase Receptor Type γ Is a Functional Tumor Suppressor Gene Specifically Downregulated in Chronic Myeloid Leukemia Marco Della Peruta1, Giovanni Martinelli4, Elisabetta Moratti1, Davide Pintani1, Marzia Vezzalini1, Andrea Mafficini1,3, Tiziana Grafone4, Ilaria Iacobucci4, Simona Soverini4, Marco Murineddu5, Fabrizio Vinante2, Cristina Tecchio2, Giovanna Piras5, Attilio Gabbas5, Maria Monne5, and Claudio Sorio1,3 Abstract Chronic myelogenous leukemia (CML) is the most common myeloproliferative disease. Protein tyrosine phosphatase receptor type γ (PTPRG) is a tumor suppressor gene and a myeloid cell marker expressed by CD34+ cells. Downregulation of PTPRG increases colony formation in the PTPRG-positive megakaryocytic cell lines MEG-01 and LAMA-84 but has no effect in the PTPRG-negative cell lines K562 and KYO-1. Its over- expression has an oncosuppressive effect in all these cell lines and is associated with myeloid differentiation and inhibition of BCR/ABL-dependent signaling. The intracellular domain of PTPRG directly interacts with BCR/ABL and CRKL, but not with signal transducers and activators of transcription 5. PTPRG is downregulated at the mRNA and protein levels in leukocytes of CML patients in both peripheral blood and bone marrow, including CD34+ cells, and is reexpressed following molecular remission of disease. Reexpression was associated with a loss of methylation of a CpG island of PTPRG promoter occurring in 55% of the patients analyzed. In K562 cell line, the DNA hypomethylating agent 5-aza-2′-deoxycytidine induced PTPRG expression and caused an inhibition of colony formation, partially reverted by downregulation of PTPRG expression. These findings establish, for the first time, PTPRG as a tumor suppressor gene involved in the pathogenesis of CML, suggesting its use as a potential diagnostic and therapeutic target. Cancer Res; 70(21); 8896–906. ©2010 AACR. Introduction For this reason, much attention has been focused on naturally occurring negative regulators of tyrosine kinase Chronic myelogenous leukemia (CML), also known as signaling: the protein tyrosine phosphatase (PTP) family chronic myeloid or chronic myelocytic leukemia, is a malig- of enzymes. nant cancer of the bone marrow myeloid lineage. It ac- The human PTP family contains 107 members, 38 of which counts for ∼15% to 20% of all cases of leukemia (1–1.5 belong to the phosphotyrosine-specific (“classic”)PTPsub- cases per 100,000 population per year; ref. 1) and originates family (subdivided in receptor- and nonreceptor-like types) from a pluripotential stem cell in which a 9:22 translocation and 61 belong to the so-called “dual-specific phosphatases.” results in the production of BCR/ABL fusion protein. This To date, only two tyrosine phosphatases, PTP1B and SHP-1, has a constitutive tyrosine kinase activity and deregulates are known to dephosphorylate and partially inhibit the trans- signal transduction pathways, leading to leukemia (2). Phos- formation potential of BCR/ABL (4, 5). Serine-threonine phorylation of key residues is required for the full transform- phosphatase PP2A is inhibited in blast crisis CML (6). These ing activity of BCR/ABL (3). PTPs belong to the nonreceptor class of enzymes. Recently, Authors' Affiliations: 1Department of Pathology and Diagnostics and performed flow cytometry. I. Iacobucci: BCR/ABL analysis. S. Soverini: 2Department of Clinical and Experimental Medicine, University of Verona; management of clinical data (Bologna). M. Murineddu: flow cytometry 3ARC-Net Research Center, University of Verona, Policlinico G.B. Rossi, and patient selection. F. Vinante: patient selection and clinical information Verona, Italy; 4Institute of Hematology and Medical Oncology, «Lorenzo (Verona). C. Tecchio: performed research. A. Mafficini: performed flow e Ariosto Seragnoli», University of Bologna, Bologna, Italy; and 5Centro cytometry. G. Piras: statistical analysis. A. Gabbas: provided clinical data. di Diagnostica Biomolecolare e Citogenetica Emato-Oncologica, “San M. Monne: patient selection and clinical information, methylation studies Francesco” Hospital, ASL3, Nuoro, Italy in patients (Nuoro). C. Sorio: designed research, wrote manuscript. Note: Supplementary data for this article are available at Cancer Research Corresponding Author: Claudio Sorio, Department of Pathology and Online (http://cancerres.aacrjournals.org/). Diagnostics, General Pathology Section, University of Verona, Strada Author contribution statement: M. Della Peruta: designed and performed Le Grazie 8, 37134 Verona, Italy. Phone: 39-45-8027688; Fax: 39-45- experiments, analyzed data. G. Martinelli: patient selection and clinical 8027127; E-mail: [email protected]. information (Bologna). E. Moratti: performed experiments, coimmunopre- doi: 10.1158/0008-5472.CAN-10-0258 cipitation studies. D. Pintani: performed clonogenic assays. M. Vezzalini: QPCR analysis, clonogenic and proliferation assays. T. Grafone: ©2010 American Association for Cancer Research. 8896 Cancer Res; 70(21) November 1, 2010 Downloaded from cancerres.aacrjournals.org on September 28, 2021. © 2010 American Association for Cancer Research. Published OnlineFirst October 19, 2010; DOI: 10.1158/0008-5472.CAN-10-0258 PTPRG in Chronic Myeloid Leukemia PTPROt, a receptor-like PTP, has been found to interfere be in complete cytogenetic remission (CCR) based on the with BCR/ABL signaling in K562 cells (7). BIOMED 2 standardized protocols. PTP receptor type γ (PTPRG) is a member of the receptor- like PTPs (8), is expressed in myeloid cells, including CD34+ Cells precursors, and can affect hematopoietic differentiation K562 (22) and MEG-01 (23) were from American Type (9–11). It is a candidate tumor suppressor gene in solid Culture Collection; KYO-1 (24) and LAMA-84 (25) were from tumors (12–14) owing to its reduced expression in ovarian, DSMZ. HEK293 was purchased from Invitrogen. K562 transfec- breast, and lung tumors (15, 16). Somatic mutations and epi- tants were selected, adding 0.50 mg/mL G418 (Invitrogen) to genetic silencing were reported (17–21). the culture medium. All were characterized by cytogenetic anal- Here, we show that PTPRG acts as a functional tumor ysis and antigen expression. Human samples were derived from suppressor gene in CML, interacting with BCR/ABL and in- Ficoll-purified cells (mRNA analysis) or whole blood samples hibiting downstream signaling events. PTPRG is specifically (flow cytometry analysis). downregulated in peripheral blood and bone marrow leuko- cytes of CML patients, at least in part by a mechanism Cell transfection and selection of CML cell lines involving hypermethylation of a PTPRG CpG island located Full-length (FL) human PTPRG (26) and antisense (AS) in the 5′ untranslated region. These findings imply that cDNA and cell lines were described (10, 27). The D1028A PTPRG might represent a potential diagnostic and thera- PTPRG cDNA was obtained by site-directed mutagenesis, peutic target in CML. wherein the Asp 1028 codon was replaced with an Ala codon (NP_002832.3) and verified by sequencing. Materials and Methods To avoid the source of error associated with clonal variation within cell lines, we selected a K562 clone (named B4) Tissue samples that maintains the capability to differentiate when treated Patients were recruited at the hematology departments of with the well-known inducers sodium butyrate and hemin Bologna, Nuoro, and Verona, Italy among newly diagnosed (28, 29) and performed all the transfection and selection CML patients during the first chronic phase. Presence of processes starting from this well-characterized clone. the Philadelphia chromosome and p210BCR-ABL rearrange- ment was a prerequisite for enrollment. CML samples were Transfection of HEK293 cells taken at diagnosis and after the initiation of therapy with HEK293 cells were transfected with FL cDNA coding for imatinib mesylate (IM). Age- and sex-matched samples p210BCR/ABL cDNA in pLNL 5LX cytomegalovirus, derived from from individuals diagnosed as not affected by malignant pLNL-XHC (30), kindly provided by Dr. Paolo Vigneri (Univer- disease were used as a reference group. Analysis of the sity of Catania, Italy). FL human PTPRG cDNA and D1028A CD34+ population for the group of patients in molecular PTPRG cDNA were previously described. Transfection was remission was not possible, as these samples were not performed with Lipofectamine 2000 reagent (Invitrogen) ac- available at the time of evaluation. Table 1 reports clinical cording to the manufacturer's instructions using 1.5 μgplasmid data. Written informed consent was obtained in accordance DNA (for single transfection) or 1 + 1 μgplasmidDNA(fordouble with the Declaration of Helsinki. Cytogenetic response was transfection) following the manufacturer's instructions. Cultures classified as complete, and the patients were considered to were cultured for 48 hours, washed with cold TBS, and lysed. Table 1. Clinical data of patients Diagnosis (n = 22) Remission (n =8) Median (range) Median (range) Age 54.12 (47–63) 60.42 (33–84) Female (%) 12.5 33.4 Bcr-Abl/Abl (%) 69.13 (30.84–100) 0.10 (0–1) Hemoglobin level (g/dL) 10.99 (7–13.8) 11.3 (5.90–15.5) Platelets (×109/L) 346.25 (6–589) 193.7 (88–291) WBC (×109/L) 218.75 (137–296) 4.77 (0.13–7.73) Peripheral blood blasts (%) 4.4 (2–6) 0 Peripheral blood basophils (%) 7.6 (0–15) 0.006 (0.002–0.01) BM blasts (%) 3.2 (1–4) 0 Palpable splenomegaly
Load more

Recommended publications
  • Identification of Chebulinic Acid As a Dual Targeting Inhibitor of Protein
    Bioorganic Chemistry 90 (2019) 103087 Contents lists available at ScienceDirect Bioorganic Chemistry journal homepage: www.elsevier.com/locate/bioorg Short communication Identification of chebulinic acid as a dual targeting inhibitor of protein T tyrosine phosphatases relevant to insulin resistance Sun-Young Yoona,1, Hyo Jin Kangb,1, Dohee Ahna, Ji Young Hwanga, Se Jeong Kwona, ⁎ Sang J. Chunga, a School of Pharmacy, Sungkyunkwan University, Suwon 16419, Republic of Korea b Department of Chemistry, Dongguk University, Seoul 100-715, Republic of Korea ARTICLE INFO ABSTRACT Keywords: Natural products as antidiabetic agents have been shown to stimulate insulin signaling via the inhibition of the Protein tyrosine phosphatases (PTPs) protein tyrosine phosphatases relevant to insulin resistance. Previously, we have identified PTPN9 and DUSP9 as Chebulinic acid potential antidiabetic targets and a multi-targeting natural product thereof. In this study, knockdown of PTPN11 Type 2 diabetes increased AMPK phosphorylation in differentiated C2C12 muscle cells by 3.8 fold, indicating that PTPN11 could Glucose-uptake be an antidiabetic target. Screening of a library of 658 natural products against PTPN9, DUSP9, or PTPN11 PTPN9 identified chebulinic acid (CA) as a strong allosteric inhibitor with a slow cooperative binding toPTPN9 PTPN11 (IC50 = 34 nM) and PTPN11 (IC50 = 37 nM), suggesting that it would be a potential antidiabetic candidate. Furthermore, CA stimulated glucose uptake and resulted in increased AMP-activated protein kinase (AMPK) phosphorylation. Taken together, we demonstrated that CA increased glucose uptake as a dual inhibitor of PTPN9 and PTPN11 through activation of the AMPK signaling pathway. These results strongly suggest that CA could be used as a potential therapeutic candidate for the treatment of type 2 diabetes.
    [Show full text]
  • PTP1B Deficiency Enables the Ability of a High-Fat Diet to Drive the Invasive Character of PTEN-Deficient Prostate Cancers
    Published OnlineFirst March 28, 2016; DOI: 10.1158/0008-5472.CAN-15-1501 Cancer Priority Report Research PTP1B Deficiency Enables the Ability of a High-Fat Diet to Drive the Invasive Character of PTEN-Deficient Prostate Cancers David P. Labbe1,2, Noriko Uetani1,Valerie Vinette1,3, Laurent Lessard4, Isabelle Aubry1, Eva Migon1, Jacinthe Sirois1, Jody J. Haigh5, Louis R. Begin 6, Lloyd C. Trotman7, Marilene Paquet8, and Michel L. Tremblay1,2,3 Abstract Diet affects the risk and progression of prostate cancer, but vation, interpreted to reflect a heightened sensitivity to IGF-1 the interplay between diet and genetic alterations in this disease stimulation upon HFD feeding. Prostate-specific overexpres- is not understood. Here we present genetic evidence in the sion of PTP1B was not sufficienttoinitiateprostatecancer, mouse showing that prostate cancer progression driven by arguingthatitactedasadiet-dependentmodifier of prostate À À loss of the tumor suppressor Pten is mainly unresponsive to cancer development in Pten / mice. Our findings offer a a high-fat diet (HFD), but that coordinate loss of the protein preclinical rationale to investigate the anticancer effects of tyrosine phosphatase Ptpn1 (encoding PTP1B) enables a highly PTP1B inhibitors currently being studied clinically for diabetes À À À À invasive disease. Prostate cancer in Pten / Ptpn1 / mice treatment as a new modality for management of prostate was characterized by increased cell proliferation and Akt acti- cancer. Cancer Res; 76(11); 3130–5. Ó2016 AACR. Introduction metabolism and cancer and is now a validated therapeutic target for diabetes, obesity, and breast cancer (7). Prostate cancer is the most frequently diagnosed cancer in The promise of PTP1B-directed therapeutics prompted us to North American men and is the second leading cause of can- further characterize the role of PTP1B in prostate cancer initiation cer-related deaths (1).
    [Show full text]
  • Inflammatory Cytokine Signalling by Protein Tyrosine Phosphatases in Pancreatic Β-Cells
    59 4 W J STANLEY and others PTPN1 and PTPN6 modulate 59: 4 325–337 Research cytokine signalling in β-cells Differential regulation of pro- inflammatory cytokine signalling by protein tyrosine phosphatases in pancreatic β-cells William J Stanley1,2, Prerak M Trivedi1,2, Andrew P Sutherland1, Helen E Thomas1,2 and Esteban N Gurzov1,2,3 Correspondence should be addressed 1 St. Vincent’s Institute of Medical Research, Melbourne, Australia to E N Gurzov 2 Department of Medicine, St. Vincent’s Hospital, The University of Melbourne, Melbourne, Australia Email 3 ULB Center for Diabetes Research, Universite Libre de Bruxelles (ULB), Brussels, Belgium esteban.gurzov@unimelb. edu.au Abstract Type 1 diabetes (T1D) is characterized by the destruction of insulin-producing β-cells Key Words by immune cells in the pancreas. Pro-inflammatory including TNF-α, IFN-γ and IL-1β f pancreatic β-cells are released in the islet during the autoimmune assault and signal in β-cells through f protein tyrosine phosphorylation cascades, resulting in pro-apoptotic gene expression and eventually phosphatases β-cell death. Protein tyrosine phosphatases (PTPs) are a family of enzymes that regulate f PTPN1 phosphorylative signalling and are associated with the development of T1D. Here, we f PTPN6 observed expression of PTPN6 and PTPN1 in human islets and islets from non-obese f cytokines diabetic (NOD) mice. To clarify the role of these PTPs in β-cells/islets, we took advantage f inflammation Journal of Molecular Endocrinology of CRISPR/Cas9 technology and pharmacological approaches to inactivate both proteins. We identify PTPN6 as a negative regulator of TNF-α-induced β-cell death, through JNK- dependent BCL-2 protein degradation.
    [Show full text]
  • The Proteintyrosine Phosphatase Receptor Type J Is Regulated by The
    Zurich Open Repository and Archive University of Zurich Main Library Strickhofstrasse 39 CH-8057 Zurich www.zora.uzh.ch Year: 2013 The Protein-Tyrosine Phosphatase Receptor Type J is regulated by the pVHL-HIF axis in Clear Cell Renal Cell Carcinoma Casagrande, Silvia ; Ruf, Melanie ; Rechsteiner, Markus ; Morra, Laura ; Brun-Schmid, Sonja ; von Teichman, Adriana ; Krek, Wilhelm ; Schraml, Peter ; Moch, Holger Abstract: Mass spectrometry analysis of renal cancer cell lines recently suggested that the Protein- Tyrosine Phosphatase Receptor Type J (PTPRJ), an important regulator of tyrosine kinase receptors, is tightly linked to the von-Hippel Lindau protein (pVHL). Therefore, we aimed to characterize the biological relevance of PTPRJ for clear cell renal cell carcinoma (ccRCC). In pVHL negative ccRCC cell lines both RNA and protein expression levels of PTPRJ were lower than those in the corresponding pVHL reconstituted cells. Quantitative RT-PCR and Western blot analysis of ccRCC with known VHL mutation status and normal matched tissues as well as RNA in situ hybridization on a Tissue Microarray (TMA) confirmed a decrease of PTPRJ expression in more than 80 % of ccRCCs, but inonly12%of papillary RCCs. ccRCC patients with no or reduced PTPRJ mRNA expression had a less favourable outcome than those with a normal expression status (p = 0.05). Sequence analysis of 32 PTPRJ mRNA negative ccRCC samples showed five known polymorphisms, but no mutations implying other mechanisms leading to PTPRJ’s down-regulation. Selective silencing of HIF-฀ by siRNA and reporter gene assays demonstrated that pVHL inactivation reduces PTPRJ expression through a HIF-dependent mechanism, which is mainly driven by HIF-2฀ stabilization.
    [Show full text]
  • The Expression Patterns and the Prognostic Roles of PTPN Family Members in Digestive Tract Cancers
    Preprint: Please note that this article has not completed peer review. The expression patterns and the prognostic roles of PTPN family members in digestive tract cancers CURRENT STATUS: UNDER REVIEW Jing Chen The First Affiliated Hospital of China Medical University Xu Zhao Liaoning Vocational College of Medicine Yuan Yuan The First Affiliated Hospital of China Medical University Jing-jing Jing The First Affiliated Hospital of China Medical University [email protected] Author ORCiD: https://orcid.org/0000-0002-9807-8089 DOI: 10.21203/rs.3.rs-19689/v1 SUBJECT AREAS Cancer Biology KEYWORDS PTPN family members, digestive tract cancers, expression, prognosis, clinical features 1 Abstract Background Non-receptor protein tyrosine phosphatases (PTPNs) are a set of enzymes involved in the tyrosyl phosphorylation. The present study intended to clarify the associations between the expression patterns of PTPN family members and the prognosis of digestive tract cancers. Method Expression profiling of PTPN family genes in digestive tract cancers were analyzed through ONCOMINE and UALCAN. Gene ontology enrichment analysis was conducted using the DAVID database. The gene–gene interaction network was performed by GeneMANIA and the protein–protein interaction (PPI) network was built using STRING portal couple with Cytoscape. Data from The Cancer Genome Atlas (TCGA) were downloaded for validation and to explore the relationship of the PTPN expression with clinicopathological parameters and survival of digestive tract cancers. Results Most PTPN family members were associated with digestive tract cancers according to Oncomine, Ualcan and TCGA data. For esophageal carcinoma (ESCA), expression of PTPN1, PTPN4 and PTPN12 were upregulated; expression of PTPN20 was associated with poor prognosis.
    [Show full text]
  • The Regulatory Roles of Phosphatases in Cancer
    Oncogene (2014) 33, 939–953 & 2014 Macmillan Publishers Limited All rights reserved 0950-9232/14 www.nature.com/onc REVIEW The regulatory roles of phosphatases in cancer J Stebbing1, LC Lit1, H Zhang, RS Darrington, O Melaiu, B Rudraraju and G Giamas The relevance of potentially reversible post-translational modifications required for controlling cellular processes in cancer is one of the most thriving arenas of cellular and molecular biology. Any alteration in the balanced equilibrium between kinases and phosphatases may result in development and progression of various diseases, including different types of cancer, though phosphatases are relatively under-studied. Loss of phosphatases such as PTEN (phosphatase and tensin homologue deleted on chromosome 10), a known tumour suppressor, across tumour types lends credence to the development of phosphatidylinositol 3--kinase inhibitors alongside the use of phosphatase expression as a biomarker, though phase 3 trial data are lacking. In this review, we give an updated report on phosphatase dysregulation linked to organ-specific malignancies. Oncogene (2014) 33, 939–953; doi:10.1038/onc.2013.80; published online 18 March 2013 Keywords: cancer; phosphatases; solid tumours GASTROINTESTINAL MALIGNANCIES abs in sera were significantly associated with poor survival in Oesophageal cancer advanced ESCC, suggesting that they may have a clinical utility in Loss of PTEN (phosphatase and tensin homologue deleted on ESCC screening and diagnosis.5 chromosome 10) expression in oesophageal cancer is frequent, Cao et al.6 investigated the role of protein tyrosine phosphatase, among other gene alterations characterizing this disease. Zhou non-receptor type 12 (PTPN12) in ESCC and showed that PTPN12 et al.1 found that overexpression of PTEN suppresses growth and protein expression is higher in normal para-cancerous tissues than induces apoptosis in oesophageal cancer cell lines, through in 20 ESCC tissues.
    [Show full text]
  • Identification of Candidate Genes and Pathways Associated with Obesity
    animals Article Identification of Candidate Genes and Pathways Associated with Obesity-Related Traits in Canines via Gene-Set Enrichment and Pathway-Based GWAS Analysis Sunirmal Sheet y, Srikanth Krishnamoorthy y , Jihye Cha, Soyoung Choi and Bong-Hwan Choi * Animal Genome & Bioinformatics, National Institute of Animal Science, RDA, Wanju 55365, Korea; [email protected] (S.S.); [email protected] (S.K.); [email protected] (J.C.); [email protected] (S.C.) * Correspondence: [email protected]; Tel.: +82-10-8143-5164 These authors contributed equally. y Received: 10 October 2020; Accepted: 6 November 2020; Published: 9 November 2020 Simple Summary: Obesity is a serious health issue and is increasing at an alarming rate in several dog breeds, but there is limited information on the genetic mechanism underlying it. Moreover, there have been very few reports on genetic markers associated with canine obesity. These studies were limited to the use of a single breed in the association study. In this study, we have performed a GWAS and supplemented it with gene-set enrichment and pathway-based analyses to identify causative loci and genes associated with canine obesity in 18 different dog breeds. From the GWAS, the significant markers associated with obesity-related traits including body weight (CACNA1B, C22orf39, U6, MYH14, PTPN2, SEH1L) and blood sugar (PRSS55, GRIK2), were identified. Furthermore, the gene-set enrichment and pathway-based analysis (GESA) highlighted five enriched pathways (Wnt signaling pathway, adherens junction, pathways in cancer, axon guidance, and insulin secretion) and seven GO terms (fat cell differentiation, calcium ion binding, cytoplasm, nucleus, phospholipid transport, central nervous system development, and cell surface) which were found to be shared among all the traits.
    [Show full text]
  • (12) Patent Application Publication (10) Pub. No.: US 2003/0082511 A1 Brown Et Al
    US 20030082511A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2003/0082511 A1 Brown et al. (43) Pub. Date: May 1, 2003 (54) IDENTIFICATION OF MODULATORY Publication Classification MOLECULES USING INDUCIBLE PROMOTERS (51) Int. Cl." ............................... C12O 1/00; C12O 1/68 (52) U.S. Cl. ..................................................... 435/4; 435/6 (76) Inventors: Steven J. Brown, San Diego, CA (US); Damien J. Dunnington, San Diego, CA (US); Imran Clark, San Diego, CA (57) ABSTRACT (US) Correspondence Address: Methods for identifying an ion channel modulator, a target David B. Waller & Associates membrane receptor modulator molecule, and other modula 5677 Oberlin Drive tory molecules are disclosed, as well as cells and vectors for Suit 214 use in those methods. A polynucleotide encoding target is San Diego, CA 92121 (US) provided in a cell under control of an inducible promoter, and candidate modulatory molecules are contacted with the (21) Appl. No.: 09/965,201 cell after induction of the promoter to ascertain whether a change in a measurable physiological parameter occurs as a (22) Filed: Sep. 25, 2001 result of the candidate modulatory molecule. Patent Application Publication May 1, 2003 Sheet 1 of 8 US 2003/0082511 A1 KCNC1 cDNA F.G. 1 Patent Application Publication May 1, 2003 Sheet 2 of 8 US 2003/0082511 A1 49 - -9 G C EH H EH N t R M h so as se W M M MP N FIG.2 Patent Application Publication May 1, 2003 Sheet 3 of 8 US 2003/0082511 A1 FG. 3 Patent Application Publication May 1, 2003 Sheet 4 of 8 US 2003/0082511 A1 KCNC1 ITREXCHO KC 150 mM KC 2000000 so 100 mM induced Uninduced Steady state O 100 200 300 400 500 600 700 Time (seconds) FIG.
    [Show full text]
  • PTPRE Sirna Set I PTPRE Sirna Set I
    Catalog # Aliquot Size P52-911B-05 3 x 5 nmol P52-911B-20 3 x 20 nmol P52-911B-50 3 x 50 nmol PTPRE siRNA Set I siRNA duplexes targeted against three exon regions Catalog # P52-911B Lot # Z2085-30 Specificity Formulation PTPRE siRNAs are designed to specifically knock-down The siRNAs are supplied as a lyophilized powder and human PTPRE expression. shipped at room temperature. Product Description Reconstitution Protocol PTPRE siRNA is a pool of three individual synthetic siRNA Briefly centrifuge the tubes (maximum RCF 4,000g) to duplexes designed to knock-down human PTPRE mRNA collect lyophilized siRNA at the bottom of the tube. expression. Each siRNA is 19-25 bases in length. The gene Resuspend the siRNA in 50 µl of DEPC-treated water accession number is BC050062. (supplied by researcher), which results in a 1x stock solution (10 µM). Gently pipet the solution 3-5 times to mix Gene Aliases and avoid the introduction of bubbles. Optional: aliquot PTPE, P-PTP-epsilon, HPTPE, R-PTP-EPSILON, RP11-380J17.1, 1x stock solutions for storage. DKFZp313F1310, FLJ57799, FLJ58245 Related Products Storage and Stability The lyophilized powder is stable for at least 4 weeks at Product Name Catalog Number room temperature. It is recommended that the PTPRA (LRP), Active P48-21G lyophilized and resuspended siRNAs are stored at or PTPRC (CD45), Active P50-21H below -20oC. After resuspension, siRNA stock solutions ≥2 PTPRC (CD45), Active P50-21BG µM can undergo up to 50 freeze-thaw cycles without PTPRE, Active P52-21G significant degradation. For long-term storage, it is PTPRF (LAR), Active P53-21G recommended that the siRNA is stored at -70oC.
    [Show full text]
  • Pdf/Infopackage Kinex.Pdf for a Com- Domain Inhibition (15)
    Protein Tyrosine Phosphatase Receptor Type γ Is a JAK Phosphatase and Negatively Regulates Leukocyte Integrin Activation This information is current as Michela Mirenda, Lara Toffali, Alessio Montresor, Giovanni of October 1, 2021. Scardoni, Claudio Sorio and Carlo Laudanna J Immunol 2015; 194:2168-2179; Prepublished online 26 January 2015; doi: 10.4049/jimmunol.1401841 http://www.jimmunol.org/content/194/5/2168 Downloaded from Supplementary http://www.jimmunol.org/content/suppl/2015/01/23/jimmunol.140184 Material 1.DCSupplemental http://www.jimmunol.org/ References This article cites 46 articles, 16 of which you can access for free at: http://www.jimmunol.org/content/194/5/2168.full#ref-list-1 Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists by guest on October 1, 2021 • Fast Publication! 4 weeks from acceptance to publication *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2015 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology Protein Tyrosine Phosphatase Receptor Type g Is a JAK Phosphatase and Negatively Regulates Leukocyte Integrin Activation Michela Mirenda,* Lara Toffali,*,† Alessio Montresor,*,† Giovanni Scardoni,† Claudio Sorio,* and Carlo Laudanna*,† Regulation of signal transduction networks depends on protein kinase and phosphatase activities.
    [Show full text]
  • A Homozygous Deletion Within the Carbonic Anhydrase-Like Domain of the Ptprg Gene in Murine L-Cells1
    (CANCER RESEARCH 53. 14«)»-1502.April1. 1993| Advances in Brief A Homozygous Deletion within the Carbonic Anhydrase-like Domain of the Ptprg Gene in Murine L-Cells1 Kishore K. Wary, Zhuangwei Lou, Arthur M. Buchberg, Linda D. Siracusa, Teresa Druck, Sal LaForgia, and Kay Huebner2 Jefferson Cancer Instilare. Thomas Jefferson Medical College, Philadelphia. Pennsylvania ¡9ÃŒ07 Abstract and murine predicted amino acid sequences for this isoform are greater than 90% identical: the Ptprg gene contains a single trans- Protein tyrosine phosphatases, on purely theoretical grounds, were sug membrane domain and two tandem tyrosine phosphatase domains. gested as possible tumor suppressor genes, and receptor protein tyrosine The extracellular region contains one fibronectin type repeat and. like phosphatase 7 (PTPRG ) has been proposed, on the basis of its location at the PTPRZ gene (7).' an NH2-terminal region of 266 amino acids with human chromosome region 3pl4.2, specifically as a tumor suppressor gene —¿30%sequence similarity to members of the carbonic anhydrase for renal cell carcinoma. We have isolated murine genomic and complementary DNA clones for enzyme family (8). analysis and mapping of the murine Ptprg locus; interspecific backcross During an investigation of the genomic organization of human and analysis showed that the Ptprg locus maps to the centramene region of mouse Ptprg loci, we noted extensive polymorphism of the murine mouse chromosome 14. We also observed a homozygous, intragenic dele Píprglocus5 and a homozygous deletion of a portion of the CA-like tion in the Ptprg gene in all donai derivatives of the original I.-cell strain, domain in murine L-cell lines, which we undertook to describe in a methylcholanthrene-treated mouse connective tissue cell line which pro detail, since homozygous deletion of a gene is one of the hallmarks of duces sarcomas in syngeneic mice.
    [Show full text]
  • Genetic Alterations of Protein Tyrosine Phosphatases in Human Cancers
    Oncogene (2015) 34, 3885–3894 © 2015 Macmillan Publishers Limited All rights reserved 0950-9232/15 www.nature.com/onc REVIEW Genetic alterations of protein tyrosine phosphatases in human cancers S Zhao1,2,3, D Sedwick3,4 and Z Wang2,3 Protein tyrosine phosphatases (PTPs) are enzymes that remove phosphate from tyrosine residues in proteins. Recent whole-exome sequencing of human cancer genomes reveals that many PTPs are frequently mutated in a variety of cancers. Among these mutated PTPs, PTP receptor T (PTPRT) appears to be the most frequently mutated PTP in human cancers. Beside PTPN11, which functions as an oncogene in leukemia, genetic and functional studies indicate that most of mutant PTPs are tumor suppressor genes. Identification of the substrates and corresponding kinases of the mutant PTPs may provide novel therapeutic targets for cancers harboring these mutant PTPs. Oncogene (2015) 34, 3885–3894; doi:10.1038/onc.2014.326; published online 29 September 2014 INTRODUCTION tyrosine/threonine-specific phosphatases. (4) Class IV PTPs include Protein tyrosine phosphorylation has a critical role in virtually all four Drosophila Eya homologs (Eya1, Eya2, Eya3 and Eya4), which human cellular processes that are involved in oncogenesis.1 can dephosphorylate both tyrosine and serine residues. Protein tyrosine phosphorylation is coordinately regulated by protein tyrosine kinases (PTKs) and protein tyrosine phosphatases 1 THE THREE-DIMENSIONAL STRUCTURE AND CATALYTIC (PTPs). Although PTKs add phosphate to tyrosine residues in MECHANISM OF PTPS proteins, PTPs remove it. Many PTKs are well-documented oncogenes.1 Recent cancer genomic studies provided compelling The three-dimensional structures of the catalytic domains of evidence that many PTPs function as tumor suppressor genes, classical PTPs (RPTPs and non-RPTPs) are extremely well because a majority of PTP mutations that have been identified in conserved.5 Even the catalytic domain structures of the dual- human cancers are loss-of-function mutations.
    [Show full text]