1 / 18

Department of Molecular Enzymology Faculty of Biotechnology, Medical University of Gdansk

Department of Molecular Enzymology Faculty of Biotechnology, Medical University of Gdansk Head: Prof. Dr Wies ł aw Makarewicz Associate Professor : Dr Andrzej C. Składanowski (see photograph) acskla@amg.gda.pl phone: (+ 48 58) 349 1470 fax: (+ 48 58) 349 1465. Project:

mbedell
Download Presentation

Department of Molecular Enzymology Faculty of Biotechnology, Medical University of Gdansk

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. Department of Molecular Enzymology Faculty of Biotechnology, Medical University of Gdansk Head: Prof. Dr Wiesław Makarewicz Associate Professor: Dr Andrzej C. Składanowski (see photograph) acskla@amg.gda.pl phone: (+ 48 58) 349 1470 fax: (+ 48 58) 349 1465

  2. Project: • Function of ecto-5’-nucleotidase in promotion adhesion and migration of melanoma cells Rafał Sądej and Andrzej C. Składanowski Current aims: • expression of ecto-5’-nucleotidase in cells • expression of structural proteins in cell membrane lipid rafts • search for potential partners of eN incell membrane • motility of cells on various stroma components

  3. Ecto-5`-nucleotidase with GPI-anchor • ,

  4. 1 2 3 4 5 6 7 Expression of ecto-5`-nucleotidase in melanoma: 1- Ab, 2- B16F10, 3- A375, 4- WM9, 5- WM35, 6- WM239, 7- WM902b Expression of N-cadherin in melanoma cells: 1- Ab, 2- B16F10, 3- A375, 4- WM9, 5- WM35,6- WM239, 7- WM902b

  5. WM9 cell lysate separated in Nycodenz gradient: gradient fractions: 2 3 4 5 6 7 8 9 K eN 2 3 4 5 6 7 8 9 α5- integrins 2 3 4 5 6 7 8 9 K α3- integrins 2 3 4 5 6 7 8 9 β3-integrin 2 3 4 5 6 7 8 9 K 200 000G 40C 4h Giα2 2 3 4 5 6 7 8 9 vimentin

  6. Izabela Rusiecka, Anna Ludwiczak, Konrad Kleszczynski, Andrzej C. Skladanowski • Być może ta prezentacja wywoła dyskusję, z której wynikną działania do wykonania. Użyj PowerPoint do śledzenia tych działań do wykonania podczas swej prezentacji • Kliknij prawym przyciskiem myszy w trakcie pokazu • Zaznacz polecenie „Notatki ze spotkania” • Zaznacz kartę „Działania do wykonania” • Wpisuj działania do wykonania w miarę ich pojawiania się • Kliknij przycisk OK, aby zamknąć okno • To automatycznie utworzy slajd działań do wykonania na końcu prezentacji wraz z punktami, które wprowadzisz. Project:OPTIMIZATION OF CELLULAR AND MOLECULAR ASSAY WITH AMP DEAMINASE FOR TOXIC AND ECO-TOXIC PROPERTIES OF VARIOUS CHEMICALS

  7. Cl- BF4- BF4- BF4- BF4- Ionic liquids Commonly used as media in the chemical industry: • dissolve both organic and inorganic compounds • show a thermal stability • have low vapour pressure • can be used in technical catalysis • are not volatile • can be recycled in technological process • [BMIM] [Cl] • [BMIM] [BF4] • [BEIM] [BF4] • [HEIM] [BF4] • [DMIM] [BF4]

  8. Contents of endogenous ammonia in ionic liquids Following concentrations of tested compounds 10-1mM 10-2 mM 10-3 mM 10-4 mM 10-5 mM 10-6 mM TBT MTO ATP CTRL CTRL+ H3PO4 [BMIM][BF4] [BMIM][Cl] [BMIM][PF6] Galaxolide [DMIM][BF4] [DMIM][Br] AMP deaminase assay on 96-well plate

  9. Project:AMP-selective 5’-nucleotidase in pigeon and human skeletal muscles Search for function of the enzyme in various muscle fibers Kinga Tkacz-Stachowska, Andrzej C. Składanowski

  10. 5’(3’)-nucleotidases of vertebrates

  11. ST 123 ST 123 Different expression of cn1a and cn1b genes in human skeletal muscles 1400 1400 1223 700 100 100 GAPDH

  12. cn1a gene is expressed on the protein level in all human skeletal muscles 43 kDa 2 3 4 5 6 7 8 9 10 11 1,3,4,5,6,8,9,10 – musculus quadriceps femoris 2- musculus brachicordialis 7- musculus tibialis anterior 11-musculus gluteus maximus

  13. 1 2 3 4 5 6 7 8 9 10 The expression of mb and cn1 genes in human skeletal muscles MyO cN-I 18S RNA

  14. Selected publications Kozłowska, M., Smoleński, R.T., Makarewicz, W., Hoffmann, C. Jastorff, B. & Świerczyński, J. (1999) ATP depletion, purine riboside triphosphate accumulation and rat thymocyte death induced by purine riboside. Toxicology Letters 104, 171-181 Sala-Newby, G.B., Składanowski, A.C. & Newby, A.C. (1999) The mechanism of adenosine formation in cells: Cloning of cytosolic 5’-nucleotidase-I. Journal of Biological Chemistry 274, 17789-17793 Minelli, A., Moroni, M., Mezzasoma, I. & Składanowski, A.C. (1999) Activity of IMP- and AMP-preferring isoforms of 5’-nucleotidase from human seminal plasma with AMP analogues. Molecular Genetics & Metabolism.66, 49-55 Tkacz, K., Cioroch, M., Składanowski, A.C., & Makarewicz, W. „The cytotoxic effect of purine riboside on COS-7 cells” in Advances in Experimental Medicine and Biology, Vol.486. Purine and Pyrimidine Metabolism in Man X (E. Zoref-Shani and Sperling O. Eds.) pp. 355-359, Kluwer Academic/Plenum Publishers; New York and London, 2000. Sala-Newby, G.B., Freeman, N.V.E., Składanowski, A.C. & Newby, A.C. (2000) Distinct roles for recombinant cytosolic 5’-nucleotidase-I and –II in AMP and IMP catabolism in COS-7 and H9c2 rat myoblast cell lines. Journal of Biological Chemistry 275, 11666-11671

  15. Future goals Long term goal: - to apply the knowledge on the function of enzymes into therapy and diagnosis. To achieve this goal we plan to set up organ and/or animal model to investigate physiological effects of application overproduced enzymes and namely cN-I into underperfused skeletal muscles. - upon revealing the complex function of ecto-5’-nucleotidase in human melanoma cells migration, we plan tosearch for diagnostic tools or therapeuticsin this type of cancer - testing AMP deaminase and other PP enzymes assays in chemical compounds risk assessment and to adopt this method in high through-put screening of compounds for commercial purposes.

More Related