Project C07 – Allosteric regulation of PAR1 biased signaling by tethered peptide ligands and receptor oligomerization
Our project will address the question how different tethered peptide agonists of protease-activated receptor 1 (PAR1) generated by thrombin or activated protein C (aPC) drive receptor signaling via different G protein or arrestin pathways. To achieve this, we will (i) determine the binding modes of thrombin- and aPC-generated tethered peptides, (ii) determine the signaling dynamics of PAR1 with G proteins and arrestins, (iii) elucidate molecular features on PAR1 and transducer proteins that impart coupling specificity and (iv) study the binding mode and molecular mode of action of parmodulins, small PAR1 biased ligands. We use experimental and simulation tools, including non-canonical amino acid-based cross-linking, mutagenesis coupled with different PAR1 functional readouts, molecular modeling and MD simulation.
Contact
Dr. Georg Künze (Project Leader)
Leipzig University, Faculty of Medicine
Junior Research Group ‘Computational Structural Biology‘
Institute for Drug Discovery
Liebigstr. 21, 04103 Leipzig
Phone +49 341 97 25726
E-Mail
Web https://www.kuenzelab.org
Shihai Jiang (PhD Student, associated)
Leipzig University Medical Center
Institute of Laboratory Medicine, Clinical Chemistry and Molecular Diagnostics
Paul-List-Straße 13/15, 04103 Leipzig
Kaja Schwerin (PhD Student)
Leipzig University, Faculty of Medicine
Junior Research Group ‘Computational Structural Biology‘
Institute for Drug Discovery
Liebigstr. 21, 04103 Leipzig
Resources
- Molecular dynamics simulations
- Molecular modeling
- In vivo models of PAR signaling
- Biochemical techniques for studying protein-protein interactions (cross linking, chimeric receptors, BRET)
Publications
Buhlan M, Ljaschenko D, Scholz N#, Langenhan T#. Experimental modulation of physiological force application on leg joint neurons in intact Drosophila melanogaster. Nature Protocols. 2024; 19:113-126. DOI: 10.1038/s41596-023-00907-7.
Manoharan J, Rana R, Kuenze G, Gupta D, Elwakiel A, Ambreen S, Wang H, Banerjee K, Zimmermann S, Singh K, Gupta A, Fatima S, Kretschmer S, Schaefer L, Zeng-Brouwers J, Schwab C, Al-Dabet MM, Gadi I, Altmann H, Koch T, Poitz DM, Baber R, Kohli S, Shahzad K, Geffers R, Lee-Kirsch MA, Kalinke U, Meiler J, Mackman N, Isermann B. Tissue factor binds to and inhibits interferon-α receptor 1 signaling. Immunity. 2024 Jan 9;57(1):68-85.e11.
Al-Dabet MM#, Shahzad K#, Elwakiel A#, Sulaj A, Kopf S, Bock F, Gadi I, Zimmermann S, Rana R, Krishnan S, Gupta D, Manoharan J, Fatima S, Nazir S, Schwab S, Baber R, Scholz M, Geffers R, Mertens PR, Nawroth PP, Griffin J, Keller M, Dockendorff C, Kohli S*, Isermann B*. Reversal of the renal hyperglycemic memory by targeting sustained tubular p21 expression. Nat Commun. 2022; 13:5062.
Madhusudhan T#, Wang H#, Straub BK, Grone E, Zhou Q, Shahzad K, Muller-Krebs S, Schwenger V, Gerlitz B, Grinnel, BW, Griffin JH, Reiser J, Grone HJ, Esmon CT, Nawroth PP, Isermann B. Cytoprotective signaling by activated protein C requires protease-activated receptor-3 in podocytes. Blood. 2012; 119:874-83.
Madhusudhan T#,*, Ghosh S#, Wang H#, Dong W, Gupta D, Elwakiel A, Stoyanov S, Al- Dabet MM, Krishnan S, Biemann R, Nazir S, Zimmermann S, Mathew A, Gadi I, Rana R, Zeng-Brouwers J, Moeller MJ, Schaefer L, Esmon CT, Kohli S, Reiser J, Rezaie AR, Ruf W, Isermann B*. Podocyte Integrin-β3 and Activated Protein C Coordinately Restrict RhoA Signaling and Ameliorate Diabetic Nephropathy. J Am Soc Nephrol. 2020; 31:1762-80.
Madhusudhan T#,*, Wang H#, Ghosh S#, Dong W, Kumar V, Al-Dabet MM, Manoharan J, Nazir S, Elwakiel A, Bock F, Kohli S, Marquardt A, Sogut I, Shahzad K, Müller AJ, Esmon CT, Nawroth PP, Reiser J, Chavakis T, Ruf W, Isermann B*. Signal integration at the PI3K-p85-XBP1 hub endows coagulation protease activated protein C with insulin-like function. Blood. 2017; 130:1445-55
Ranjan S, Goihl S, Kohli S, Gadi I, Piera M, Shahzad K, Gupta D, Bock F, Wang H, Shaik H, Kähne T, Reinhold D, Bank U, Zenclussen AC, Niemz J, Schnöder TM, Brunner-Weinzierl M, Fischer T, Kalinski T, Schraven B, Luft T, Huehn J, Naumann M, Heidel FH, Isermann B. Activated protein C protects from GvHD via PAR2/PAR3 signalling in regulatory T-cells. Nat Commun. 2017; 8:311.
Marlow B, Kuenze G, Li B, Sanders CR, Meiler J. Structural determinants of cholesterol recognition in helical integral membrane proteins. Biophys J. 2021; 120:1592-1604.
Kuenze G∆, Vanoye CG, Desai RR, Adusumilli S, Brewer KR, Woods H, McDonald, EF, George AL Jr, Sanders CR, Meiler J. Allosteric Mechanism for KCNE1 Modulation of KCNQ1 Potassium Channel Activation. eLife 2020; 9:e57680. ∆first and corresponding author.
Künze G, Isermann B*. Targeting biased signaling by PAR1: Function and molecular mechanism of parmodulins. Blood. 2023; Accepted for publication.
O9. Kuenze G*, Bonneau R, Koehler Leman J*, Meiler J*. Integrative protein modeling in RosettaNMR from sparse paramagnetic restraints. Structure. 2019; 27:1721-34.
Jödicke L#, Mao J#, Kuenze G#, Reinhart C, Kalavacherla T, Jonker HRA, Richter C, Schwalbe H, Meiler J, Preu J, Michel H*, Glaubitz C*. The molecular basis of subtype selectivity of human kinin G-protein-coupled receptors. Nat Chem Biol. 2018; 14:284-90.
