Title:
APPL PROTEINS AS RAB5 EFFECTORS
Document Type and Number:
WIPO Patent Application WO2005005475
Kind Code:
A3
Abstract:
The present invention relates to an in vivo-assay to screen for anti-proliferative drugs, the assay comprising the steps of: (a) contacting cells of a primary cell culture or of an established cell line with a candidate substance, (b) subsequently or concomitantly with a candidate substance, contacting the cells with a growth factor, (c) processing the cells for immunofluorescence staining to detect APPL1 and APPL2 using an anti-APPL1 and/or 2 antibody, or alternatively using GFP-tagged APPL proteins stably or transiently expressed by the cells via transfection, (d) assessing the degree of colocalisation of APPL1 and/or 2 and the growth factor, the solubilisation of APPL1 and/or 2 and their translocation to the nucleus, (e) repeating steps (b) to (d) with cells not previously treated with the candidate substance, and (f) comparing the degree of colocalisation of APPL1 and/or 2 and the growth factor, the solubilisation of APPL1 and/or 2 and their translocation to the nucleus between the cells not previously treated with the candidate substance (untreated cells) and cells treated with the candidate substance (treated cells), wherein an altered degree of colocalisation of APPL1 and/or 2 and the growth factor, an altered solubilisation of APPL1 and/or 2 and/or their altered translocation to the nucleus in the treated vs. the untreated cells identifies the candidate substance as an anti-proliferative drug.
Inventors:
ZERIAL MARINO (DE)
MIACZYNSKA MARTA (DE)
MIACZYNSKA MARTA (DE)
Application Number:
PCT/EP2004/007527
Publication Date:
May 26, 2005
Filing Date:
July 08, 2004
Export Citation:
Assignee:
MAX PLANCK GESELLSCHAFT (DE)
ZERIAL MARINO (DE)
MIACZYNSKA MARTA (DE)
ZERIAL MARINO (DE)
MIACZYNSKA MARTA (DE)
International Classes:
C07K14/47; (IPC1-7): A61P35/00; A61K45/00
Domestic Patent References:
| WO2001020022A1 | 2001-03-22 |
Foreign References:
| EP1088898A1 | 2001-04-04 |
Other References:
MIACZYNSKA MARTA ET AL: "APPL proteins link Rab5 to nuclear signal transduction via an endosomal compartment.", CELL, vol. 116, no. 3, 6 February 2004 (2004-02-06), pages 445 - 456, XP002318580, ISSN: 0092-8674
HABERMANN BIANCA: "The BAR-domain family of proteins: a case of bending and binding?", EMBO REPORTS. MAR 2004, vol. 5, no. 3, March 2004 (2004-03-01), pages 250 - 255, XP002318581, ISSN: 1469-221X
MIACZYNSKA MARTA ET AL: "Not just a sink: endosomes in control of signal transduction.", CURRENT OPINION IN CELL BIOLOGY. AUG 2004, vol. 16, no. 4, June 2004 (2004-06-01), pages 400 - 406, XP002318582, ISSN: 0955-0674
DATABASE EMBL [online] 27 June 2002 (2002-06-27), "Homo sapiens DIP13 beta mRNA, complete cds.", XP002318584, retrieved from EBI accession no. EM_PRO:AY113704 Database accession no. AY113704
MITSUUCHI Y ET AL: "Identification of a chromosome 3p14.3-21.1 gene, APPL, encoding an adaptor molecule that interacts with the oncoprotein-serine/threonine kinase AKT2", ONCOGENE, BASINGSTOKE, HANTS, GB, vol. 18, 1999, pages 4891 - 4898, XP002965023, ISSN: 0950-9232
DE RENZIS STEFANO ET AL 00022002: "Divalent Rab effectors regulate the sub-compartmental organization and sorting of early endosomes", NATURE CELL BIOLOGY, vol. 4, no. 2, February 2002 (2002-02-01), pages 124 - 133, XP002318583, ISSN: 1465-7392
HABERMANN BIANCA: "The BAR-domain family of proteins: a case of bending and binding?", EMBO REPORTS. MAR 2004, vol. 5, no. 3, March 2004 (2004-03-01), pages 250 - 255, XP002318581, ISSN: 1469-221X
MIACZYNSKA MARTA ET AL: "Not just a sink: endosomes in control of signal transduction.", CURRENT OPINION IN CELL BIOLOGY. AUG 2004, vol. 16, no. 4, June 2004 (2004-06-01), pages 400 - 406, XP002318582, ISSN: 0955-0674
DATABASE EMBL [online] 27 June 2002 (2002-06-27), "Homo sapiens DIP13 beta mRNA, complete cds.", XP002318584, retrieved from EBI accession no. EM_PRO:AY113704 Database accession no. AY113704
MITSUUCHI Y ET AL: "Identification of a chromosome 3p14.3-21.1 gene, APPL, encoding an adaptor molecule that interacts with the oncoprotein-serine/threonine kinase AKT2", ONCOGENE, BASINGSTOKE, HANTS, GB, vol. 18, 1999, pages 4891 - 4898, XP002965023, ISSN: 0950-9232
DE RENZIS STEFANO ET AL 00022002: "Divalent Rab effectors regulate the sub-compartmental organization and sorting of early endosomes", NATURE CELL BIOLOGY, vol. 4, no. 2, February 2002 (2002-02-01), pages 124 - 133, XP002318583, ISSN: 1465-7392
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