[go: up one dir, main page]

CA2277430A1 - Systeme d'expression stable d'une phosphodiesterase de camp a affinite elevee et son utilisation - Google Patents

Systeme d'expression stable d'une phosphodiesterase de camp a affinite elevee et son utilisation Download PDF

Info

Publication number
CA2277430A1
CA2277430A1 CA002277430A CA2277430A CA2277430A1 CA 2277430 A1 CA2277430 A1 CA 2277430A1 CA 002277430 A CA002277430 A CA 002277430A CA 2277430 A CA2277430 A CA 2277430A CA 2277430 A1 CA2277430 A1 CA 2277430A1
Authority
CA
Canada
Prior art keywords
enzyme
pde
rolipram
camp
iva
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Abandoned
Application number
CA002277430A
Other languages
English (en)
Inventor
Douglas J. Pon
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Merck Frosst Canada and Co
Original Assignee
Individual
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from GBGB9705910.9A external-priority patent/GB9705910D0/en
Application filed by Individual filed Critical Individual
Publication of CA2277430A1 publication Critical patent/CA2277430A1/fr
Abandoned legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/34—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase
    • C12Q1/44—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase involving esterase
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/531—Production of immunochemical test materials
    • G01N33/532—Production of labelled immunochemicals
    • G01N33/535—Production of labelled immunochemicals with enzyme label or co-enzymes, co-factors, enzyme inhibitors or enzyme substrates

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Immunology (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • Molecular Biology (AREA)
  • Zoology (AREA)
  • General Health & Medical Sciences (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Microbiology (AREA)
  • Urology & Nephrology (AREA)
  • Analytical Chemistry (AREA)
  • Wood Science & Technology (AREA)
  • Physics & Mathematics (AREA)
  • Biochemistry (AREA)
  • Hematology (AREA)
  • Biomedical Technology (AREA)
  • Biotechnology (AREA)
  • Genetics & Genomics (AREA)
  • General Engineering & Computer Science (AREA)
  • Cell Biology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biophysics (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Enzymes And Modification Thereof (AREA)

Abstract

On a déterminé une lignée cellulaire CHO-K1 exprimant de façon stable une enzyme humaine PDE IVa de recombinaison à longueur totale (rhPDEIVa) par sélection d'hygromycine B. On a évalué l'inhibition de l'activité de PDE IVa exprimée par des inhibiteurs sélectifs de PDE IV. L'ordre de classification des puissances des inhibiteurs en élevant cAMP dans la méthode de détermination de cellules entières s'est avéré totalement différent de celui obtenu avec l'enzyme soluble. L'ordre de classification des puissances s'est également maintenu quand on a mesuré des rapports d'activité de PKA à la place de niveaux de cAMP. Quand on a examiné l'inhibition de l'activité de PDE IVa soluble en présence de 100 mM de MgCl¿2?, l'ordre de classification des puissances des inhibiteurs s'est avéré être identique à celui qu'on avait obtenu pour les méthodes de détermination de cellules entières (à la fois cAMP et PLA). La conformation observée d'affinité élevée de l'enzyme ne dépendait pas d'un cation ou d'un anion spécifique. Le déplacement de sensibilité de l'enzyme rhPDE IVa pour (R)-rolipram en présence d'une force ionique accrue était accompagné par une capacité améliorée de la préparation d'enzyme soluble à se fixer de façon spécifique et avec une affinité élevée à [3H](R)-rolipram.
CA002277430A 1997-01-09 1998-01-07 Systeme d'expression stable d'une phosphodiesterase de camp a affinite elevee et son utilisation Abandoned CA2277430A1 (fr)

Applications Claiming Priority (7)

Application Number Priority Date Filing Date Title
US3469197P 1997-01-09 1997-01-09
US60/034,691 1997-01-09
GBGB9705910.9A GB9705910D0 (en) 1997-03-21 1997-03-21 System for stably expressing a high-affinity cAMP phosphodiesterase and use thereof
GB9705910.9 1997-03-21
US6542097P 1997-11-13 1997-11-13
US60/065,420 1997-11-13
PCT/CA1998/000024 WO1998030718A1 (fr) 1997-01-09 1998-01-07 Systeme d'expression stable d'une phosphodiesterase de camp a affinite elevee et son utilisation

Publications (1)

Publication Number Publication Date
CA2277430A1 true CA2277430A1 (fr) 1998-07-16

Family

ID=27268783

Family Applications (1)

Application Number Title Priority Date Filing Date
CA002277430A Abandoned CA2277430A1 (fr) 1997-01-09 1998-01-07 Systeme d'expression stable d'une phosphodiesterase de camp a affinite elevee et son utilisation

Country Status (2)

Country Link
CA (1) CA2277430A1 (fr)
WO (1) WO1998030718A1 (fr)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6368815B1 (en) * 1999-03-29 2002-04-09 Warner-Lambert Company Screening of molecules that inhibit human phosphodiesterase 4A produced by non-recombinant cell lines

Family Cites Families (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
TW263495B (fr) * 1992-12-23 1995-11-21 Celltech Ltd
DE69434492T2 (de) * 1993-03-10 2006-07-13 Smithkline Beecham Corp. Phosphodiesterase des menschlichen gehirns
DE69433997T2 (de) * 1993-06-18 2005-01-27 Smithkline Beecham Corp. Verfahren zur Identifizierung eines PDE IV Hemmers
GB9412571D0 (en) * 1994-06-22 1994-08-10 Celltech Ltd Chemical compounds
GB2301363B (en) * 1994-12-23 1998-12-30 Celltech Therapeutics Ltd Human phosphodiesterase type ivc and its production and use
GB9603723D0 (en) * 1996-02-22 1996-04-24 Merck & Co Inc Diphenyl pyridyl derivatives as pde iv inhibitors

Also Published As

Publication number Publication date
WO1998030718A1 (fr) 1998-07-16

Similar Documents

Publication Publication Date Title
Bolger et al. The unique amino-terminal region of the PDE4D5 cAMP phosphodiesterase isoform confers preferential interaction with β-arrestins
Yarwood et al. The RACK1 signaling scaffold protein selectively interacts with the cAMP-specific phosphodiesterase PDE4D5 isoform
Bamette et al. Association of the anti-inflammatory activity of phosphodiesterase 4 (PDE4) inhibitors with either inhibition of PDE4 catalytic activity or competition for [3H] rolipram binding
Kawabe et al. Differential activation of adenylyl cyclase by protein kinase C isoenzymes
Gagnon et al. Characterization of SPAK and OSR1, regulatory kinases of the Na-K-2Cl cotransporter
Vullo et al. Carbonic anhydrase inhibitors. Inhibition of the transmembrane isozyme XII with sulfonamides—a new target for the design of antitumor and antiglaucoma drugs?
Krugmann et al. Characterizing the interactions between the two subunits of the p101/p110γ phosphoinositide 3-kinase and their role in the activation of this enzyme by Gβγ subunits
MACKENZIE et al. Action of rolipram on specific PDE4 cAMP phosphodiesterase isoforms and on the phosphorylation of cAMP-response-element-binding protein (CREB) and p38 mitogen-activated protein (MAP) kinase in U937 monocytic cells
Saldou et al. Comparison of recombinant human PDE4 isoforms: interaction with substrate and inhibitors
US5922557A (en) System for stably expressing a high-affinity camp phosphodiesterase and use thereof
Bajpai et al. AKAP3 selectively binds PDE4A isoforms in bovine spermatozoa
Shepherd et al. Molecular cloning and subcellular distribution of the novel PDE4B4 cAMP-specific phosphodiesterase isoform
Ming et al. The Rac effector p67 phox regulates phagocyte NADPH oxidase by stimulating Vav1 guanine nucleotide exchange activity
MacKenzie et al. Phosphorylation of cAMP-specific PDE4A5 (phosphodiesterase-4A5) by MK2 (MAPKAPK2) attenuates its activation through protein kinase A phosphorylation
Van Hoof et al. Specific interactions of PP2A and PP2A-like phosphatases with the yeast PTPA homologues, Ypa1 and Ypa2
Perrino et al. Substrate selectivity and sensitivity to inhibition by FK506 and cyclosporin A of calcineurin heterodimers composed of the α or β catalytic subunit
Walker et al. Protein kinase C chimeras: catalytic domains of alpha and beta II protein kinase C contain determinants for isotype-specific function.
Laffargue et al. Phosphoinositide 3‐kinase and integrin signalling are involved in activation of Bruton tyrosine kinase in thrombin‐stimulated platelets
POOLEY et al. Intracellular localization of the PDE4A cAMP-specific phosphodiesterase splice variant RD1 (RNPDE4A1A) in stably transfected human thyroid carcinoma FTC cell lines
US6329137B1 (en) Intact cell assay for protein tyrosine phosphatases using recombinant baculoviruses
Kahlos et al. Thioredoxin restores nitric oxide-induced inhibition of protein kinase C activity in lung endothelial cells
Ireton et al. Expression of human topoisomerase I with a partial deletion of the linker region yields monomeric and dimeric enzymes that respond differently to camptothecin
Weeks et al. N-Terminal domain of phosphodiesterase-11A4 (PDE11A4) decreases affinity of the catalytic site for substrates and tadalafil, and is involved in oligomerization
Owens et al. Molecular cloning and expression of a human phosphodiesterase 4C
EP0889971B1 (fr) Modulateur de la phosphatase

Legal Events

Date Code Title Description
EEER Examination request
FZDE Discontinued