Struktur-Funktionsbeziehungen im funktionellen Komplex der membranständigen neuronalen Guanylatzyklase aus Photorezeptoren

• Applicants: Professor Dr. Alfred Blume; Professor Dr. Daniel Huster; Professorin Dr. Andrea Sinz
• Subject Area: Biophysics
• Term: from 2009 to 2015
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 131296762

• Final Report Year: 2015

Final Report Abstract

In the course of this project, we were able to derive first structural models of myristoylated GCAP-2 and a GCAP-2/GC peptide complex confirming the postulated binding site between ROS-GC and GCAP-2. Also, the first structural model was presented for the GCAP-2 homodimer. Chemical cross-linking using stable isotope (15N) labeled protein was found to be indispensable for discriminating between intra- and intermolecular cross-links, i.e., within one monomer or between different monomers.

Publications

• StavroX – A Software for Analyzing Cross-Linked Products in Protein Interaction Studies, J. Am. Soc. Mass Spectrom. 23, 76-87 (2012)
  Götze, Michael; Pettelkau, Jens; Schaks, Sabine; Bosse, Konstanze; Ihling, Christian H.; Krauth, Fabian; Fritzsche, Romy; Kühn, Uwe & Sinz, Andrea
  
• The interaction of lipid modified pseudopeptides with lipid membranes, Org. Biomol. Chem. 9, 6998-7006 (2011)
  Scheidt, A., Sickert, A., Meier, T., Castellucci, N., Tomasini, C., Huster, D.
  
• The Presence of Membranes or Micelles Induces Structural Changes of the Myristoylated Guanylate-Cyclase Activating Protein-2, Eur. Biophys. J. 40 565-576 (2011)
  Theisgen, S., Thomas, L., Schröder, T., Lange, C., Kovermann, M., Balbach, J., Huster, D.
  
• Structural Insights into Retinal Guanylylcyclase / GCAP-2 Interaction by Cross-Linking and Mass Spectrometry, Biochemistry 51, 4932-4949 (2012)
  Pettelkau, J., Schröder, T., Ihling, C.H., Olausson, B.E., Kölbel, K., Lange, C., Sinz, A.
  
• Peptide and protein binding to lipid monolayers studied by FT-IRRA spectroscopy, Biochim. Biophys. Acta (Biomembranes) 1828, 2294-2305 (2013)
  Blume, Alfred & Kerth, Andreas
  
• Structural analysis of guanylyl cyclase-activating protein-2 (GCAP-2) homodimer by stable isotope-labeling, chemical cross-linking, and mass spectrometry, J. Am. Soc. Mass Spectrom. 24, 1969-79 (2013)
  Pettelkau, Jens; Thondorf, Iris; Theisgen, Stephan; Lilie, Hauke; Schröder, Thomas; Arlt, Christian; Ihling, Christian H. & Sinz, Andrea
  
• Accessibilities of N-terminal myristoyl chain and cysteines in guanylyl cyclase-activating protein-2 (GCAP-2) studied by covalent labeling and mass spectrometry. Rapid Commun. Mass Spectrom. 28, 835-838 (2014)
  Ihling, Christian H.; Schröder, Thomas; Pettelkau, Jens; Tischer, Alexander; Lange, Christian & Sinz, Andrea
  
• Automated Assignment of MS/MS Cleavable Cross-Links in Protein 3D-Structure Analysis, J. Am. Soc. Mass Spectrom. 26, 83-97 (2015)
  Götze, Michael; Pettelkau, Jens; Fritzsche, Romy; Ihling, Christian H.; Schäfer, Mathias & Sinz, Andrea
  
• Guanylate cyclase-activating protein-2 undergoes structural changes upon binding to lipid surfaces, Biochim. Biophys. Acta (Biomembranes) 1838, 2767-2777 (2014)
  Margetić, Aleksandra; Nannemann, David; Meiler, Jens; Huster, Daniel & Theisgen, Stephan
  
• Reliable Identification of Cross-Linked Products in Protein Interaction Studies by 13C-Labeled p-Benzoylphenylalanine, J. Am. Soc. Mass Spectrom. 25, 1628-1641 (2014)
  Pettelkau, Jens; Ihling, Christian H.; Frohberg, Petra; van Werven, Lars; Jahn, Olaf & Sinz, Andrea