A05

A05: Summary

In this project the activation of the ion channel TRPM2 by the nucleotides ADPR and 2dADPR will be investigated by visualization of channel activity, degradation of the ligands and specific antagonists. In addition, a structural model for the intact tetrameric TRPM2 channel will be developed by cryo-electron microscopy, that, in conjunction with a high-resolution crystal structure of the nucleotide-binding domain of TRPM2 in complex with a regulatory nucleotide will lead to a detailed understanding of the gating mechanism and the different effects of the physiological ligands.

Dr. Ralf Fliegert

Selected publications

 

  1. Fliegert R, Riekehr WM, Guse AH (2020) Does Cyclic ADP-Ribose (cADPR) Activate the Non-selective Cation Channel TRPM2?. Front Immunol 11:2018
  2. Baszczyňski O, Watt JM, Rozewitz MD, Guse AH, Fliegert R, Potter BVL (2019) Synthesis of Terminal Ribose Analogues of Adenosine 5'-Diphosphate Ribose as Probes for the Transient Receptor Potential Cation Channel TRPM2. J Org Chem  84:6143-6157
  3. Fliegert R, Bauche A, Wolf Pérez AM, Watt JM, Rozewitz MD, Winzer R, Janus M, Gu F, Rosche A, Harneit A, Flato M, Moreau C, Kirchberger T, Wolters V, Potter BVL, Guse AH (2017) 2'-Deoxyadenosine 5'-diphosphoribose is an endogenous TRPM2 superagonist. Nat Chem Biol 13:10361044.
  4. Fliegert R, Watt JM, Schöbel A, Rozewitz MD, Moreau C, Kirchberger T, Thomas MP, Sick W, Araujo AC, Harneit A, Potter BVL, Guse AH (2017) Ligand-induced activation of human TRPM2 requires the terminal ribose of ADPR and involves Arg1433 and Tyr1349. Biochem J 474:2159-2175.
  5. Wolf IM, Diercks BP, Gattkowski E, Czarniak F, Kempski J, Werner R, Schetelig D, Mittrücker HW, Schumacher V, von Osten M, Lodygin D, Flügel A, Fliegert R, Guse AH (2015) Frontrunners of T cell activation: Initial, localized Ca2+ signals mediated by NAADP and the type 1 ryanodine receptor. Sci Signal 8:ra102.
  6. Guse AH, Ernst IM, Fliegert R (2013) NAADP signaling revisited. Curr Top Med Chem 13:29782990.
  7. Moreau C, Kirchberger T, Swarbrick JM, Bartlett SJ, Fliegert R, Yorgan T, Bauche A, Harneit A, Guse AH, Potter BV (2013) Structure-activity relationship of adenosine 5'-diphosphoribose at the transient receptor potential melastatin 2 (TRPM2) channel: rational design of antagonists. J Med Chem 56:10079-10102.
  8. Schwarz N*, Fliegert R*, Adriouch S*, Seman M, Guse AH, Haag F, Koch-Nolte F (2009) Activation of the P2X7 ion channel by soluble and covalently bound ligands. Purinergic Signal 5:139-149. *equal contribution as first authors
  9. Fliegert R, Gasser A, Guse AH (2007) Regulation of calcium signalling by adenine-based second messengers. Biochem Soc Trans 35:109-114.
  10. Fliegert R, Glassmeier G, Schmid F, Cornils K, Genisyuerek S, Harneit A, Schwarz JR, Guse AH (2007) Modulation of Ca2+ entry and plasma membrane potential by human TRPM4b. FEBS J 274:704713.

Prof. Dr. Henning Tidow

Selected publications

 

  1. Gattkowski E, Johnsen A, Bauche A, Möckl F, Kulow F, Garcia Alai M, Rutherford TJ, Fliegert R, Tidow H (2019) Novel CaM-binding motif in its NudT9H domain contributes to temperature sensitivity of TRPM2. Biochim Biophys Acta Mol Cell Res 1866:1162-1170
  2. Veith K, Molledo MM, Hernandez YA, Josts I, Nitsche J, Löw C, Tidow H (2017) Lipid-like peptides can stabilize integral membrane proteins for biophysical and structural studies. Chembiochem 18:17351742.
  3. Josts I, Almeida Hernandez Y, Andreeva A, Tidow H (2016) Crystal Structure of a Group I Energy Coupling Factor Vitamin Transporter S Component in Complex with Its Cognate Substrate. Cell Chem Biol 23:827-836.
  4. Tidow H, Nissen P (2013) Structural diversity of CaM-binding to its target sites. FEBS J 280:55515565.
  5. Tidow H, Poulsen LR, Andreeva A, Knudsen M, Hein KL, Wiuf C, Palmgren MB, Nissen P (2012) A bimodular mechanism of calcium control in eukaryotes. Nature 491:468-472.
  6. Tidow H, Andreeva, A, Rutherford, TJ, Fersht, AR (2009) Solution structure of U11-48K CHHC zinc finger domain that specifically binds the 5’ splice site of U12-type introns. Structure 17:294-302.
  7. Andreeva A, Tidow H (2008) A novel CHHC Zn-finger domain found in spliceosomal proteins and tRNA modifying enzymes. Bioinformatics 24:2277-2280.
  8. Wells M, Tidow H, Rutherford TJ, Markwick P, Jensen MR, Mylonas E, Svergun DI, Blackledge M, Fersht AR (2008) Structure of tumor suppressor p53 and its intrinsically disordered N-terminal transactivation domain. Proc Natl Acad Sci USA 105:5762-5767.
  9. Tidow H, Melero R, Mylonas E, Freund SM, Grossmann, JG, Carazo KM., Svergun DI, Valle M, Fersht AR (2007) Quaternary structures of tumor suppressor p53 and a specific p53 DNA complex. Proc Natl Acad Sci USA 104: 12324-12329.
  10. Tidow H, Andreeva A., Rutherford TJ, Fersht AR (2007) Solution Structure of ASPP2 N-terminal Domain (N-ASPP2) Reveals a Ubiquitin-like Fold. J Mol Biol 371:948-958.

Our Team

Prof. Dr. Henning Tidow

Dept. of Chemistry & The Hamburg Centre for Ultrafast Imaging (CUI)
 
University of Hamburg

Dr. Ralf Fliegert

Department of Biochemistry and Molecular Cell Biology

University Medical Centre Hamburg-Eppendorf (UKE)

M.Sc. Winnie Riekehr

Department of Biochemistry and Molecular Cell Biology

University Medical Centre Hamburg-Eppendorf (UKE)

M.Sc. Ellen Gattkowski

Dept. of Chemistry & The Hamburg Centre for Ultrafast Imaging (CUI)
 
University of Hamburg

M.Sc. Simon Sander

Dept. of Chemistry & The Hamburg Centre for Ultrafast Imaging (CUI)
 
University of Hamburg

Jelena Pick

Department of Biochemistry and Molecular Cell Biology

University Medical Center Hamburg-Eppendorf (UKE)

Contact

University Medical Center Hamburg-Eppendorf 
Department of Biochemistry and Molecular Cell Biology 
Martinistrasse 52
20246 Hamburg

Scientific Coordinator

Dr. Björn-Philipp Diercks
Fon: +49  (0) 40 7410 54338
E-Mail: b.diercks©uke.de

Administration

Laura Mitsching
Fon: +49  (0) 40 7410 50301
E-Mail: l.mitsching©uke.de