A05: SFB 1328

A05 Project movie

A05

TRPM2 is a non-selective, Ca2+-permeable ion channel expressed in immune cells that can be activated by ADPR and 2dADPR. Within this project we will analyse the role of two distinct nucleotide binding domains for channel activation and their interplay with a recently identified Zn2+ binding site, and solve the structure of TRPM2 in its native lipid environment and in complex with PI(4,5)P2 and calmodulin to improve our understanding of the pleiotropic regulation of the channel. In addition we will investigate the spatio-temporal organization of the ADPR/2dADPR signalling pathway during chemotaxis of neutrophil granulocytes.

Dr. Ralf Fliegert

E-Mail: r.fliegert©uke.uni-hamburg.de
www: Dept. of Biochemistry and Molecular Cell Biology, University Medical Centre HamburgEppendorf (UKE)

Maria Marta Garcia Alai,PhD

E-Mail: garcia©embl-hamburg.de
www: Integrative structural biology and Molecular biophysics & European Molecular Biology Laboratory (EMBL), Centre for Structural Systems Biology (CSSB)

Dr. Ralf Fliegert

Selected publications (*equal contribution)

Fliegert R, Riekehr WM, Guse AH (2020) Does Cyclic ADP-Ribose (cADPR) Activate the Non-selective Cation Channel TRPM2?. Front Immunol 11:2018
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
Fliegert R*, Heeren J*, Koch-Nolte F*, Nikolaev VO*, Lohr C*, Meier C*, Guse AH* (2019) Adenine nucleotides as paracrine mediators and intracellular second messengers in immunity and inflammation. Biochem Soc Trans 47(1):329-337.
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 66(7):1162-1170.
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.
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.
Guse AH, Ernst IM, Fliegert R (2013) NAADP signaling revisited. Curr Top Med Chem 13:29782990.
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.
Fliegert R, Gasser A, Guse AH (2007) Regulation of calcium signalling by adenine-based second messengers. Biochem Soc Trans 35:109-114.
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

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
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.
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.
Tidow H, Nissen P (2013) Structural diversity of CaM-binding to its target sites. FEBS J 280:55515565.
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.
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.
Andreeva A, Tidow H (2008) A novel CHHC Zn-finger domain found in spliceosomal proteins and tRNA modifying enzymes. Bioinformatics 24:2277-2280.
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.
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.
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.