Signaling mechanism, subcellular localization and regulation of guanylyl cyclases A and B, the receptors for atrial and C-type natriuretic peptides

Project Description
Principal Investigator(s)

The natriuretic peptides (NPs) ANP, BNP and CNP have diverse physiological, i.e. cardiovascular actions. Their cyclic GMP forming guanylyl cyclase (GC) receptors, GC-A (shared by ANP and BNP) and GC-B (for CNP) are members of a family of seven particulate (p)GCs receptors (A to G) with different ligands and functions. They all exist as homodimers of single-span transmembrane proteins, containing an extracellular ligand-binding domain (ECD) and three intracellular domains, including the C-terminal guanylyl cyclase domain (GCD). The mechanism of ligand-dependent activation of cGMP production by pGC receptors is ultimately unknown. The crystal structures of the GC-A-ECD dimer and the ANP-ECD complex (1:2) suggested that ligand-induced rotation of the ECDs is transmitted across the membrane and reorients the two intracellular GC domains into the active conformation, thereby enabling cGMP synthesis.

In this project we aim to visualize the dynamics of ligand-induced conformational changes of GC-A (as representative member of the family of pGC receptors) in living cells by inserting genetically encoded fluorescent tags. We will use a FRET-FlAsH approach to study the ligand-evoked conformational changes of GC-A and the impact of receptor dephosphorylation/desensitization. In addition we plan to use single-molecule microscopy and super-resolution imaging (with dSTORM) to directly observe the distinct subcellular localizations of GC-A and GC-B in cardiomyocytes, the role of receptor internalization in GC-A/GC-B desensitization, and the subcellular (re)distribution of these receptors in cardiomyocyte hypertrophy.

Title Year Authors Journal Links
Atrial Natriuretic Peptide Locally Counteracts the Deleterious Effects of Cardiomyocyte Mineralcorticoid Receptor Activation 2014 Nakagawa, H., Oberwinkler, H., Nikolaev, V. O., Gaßner, B., Umbenhauer, S., Wagner, H., Saito, Y., Baba, H. A., Frantz, S., and Kuhn, M. Circ Heart Fail More
Transgenic Mice for Real Time Visualization of cGMP in Intact Adult Cardiomyocytes 2014 Götz, K., Sprenger, J., Perera, R. K., Steinbrecher, J. H., Lehnart, S. E., Kuhn, M., Gorelik, J., Balligand, J. L., and Nikolaev, V. O . Circ Res More
Stress - dependent dilated cardiomyopathy in mice with cardiomyocyte - restricted inactivation of cyclic GMP-dependent protein kinase I 2013 Frantz, S., Klaiber, M., Baba, H. A., Oberwinkler, H., Völker, K., Gaßner, B., Feil, R., Hofmann, F., and Kuhn, M. Eur Heart J More
An activating mutation in the kinase homology domain of the natriuretic peptide receptor-2 causes extremely tall stature without skeletal deformities 2013 Hannema S. E., van Duyvenvoorde, H. A., Premsler, T., Yang, R.-B., Müller, T. D., Gaßner, B., Oberwinkler, H., Pereira, A. M., Losekoot, M., Kuhn, M.*, and Wit, J. M.* J Clin Endocrinol Metab More
A cardiac pathway of cyclic GMP-independent signaling of guanylyl cyclase A, the receptor for atrial natriuretic peptide 2011 Klaiber, M., Dankworth, B., Kruse, M., Hartmann, M., Nikolaev, V. O., Yang, R. B., Völker, K., Gaßner, B., Oberwinkler, H., Feil, R., Freichel, M., Groschner, K., Skryabin, B. V., Frantz, S., Birnbaumer, L., Pongs, O., and Kuhn, M. Proc Natl Acad Sci USA More
Homologous desensitization of guanylyl cyclase A, the receptor for atrial natriuretic peptide, is associated with a complex phosphorylation pattern 2010 Schröter, J., Zahedi, R. P., Hartmann, M., Gassner, B., Gazinski, A., Waschke, J., Sickmann, A., and Kuhn, M. FEBS J More
The natriuretic peptide / guanylyl cyclase-A system functions as a stress-responsive regulator of angiogenesis in mice 2009 Kuhn, M., Völker, K., Schwarz, K., Flögel, U., Jacoby, C., Stypmann, J., van Eickels, M., Gambaryan, S., Werner, M., Hartmann, M., Schrader, J., and Baba H. A. J Clin Invest More
Vascular endothelium is critically involved in the hypotensive and hypovolemic actions of atrial natriuretic peptide 2005 Sabrane, K., Kruse, M. N., Fabritz, L., Zwiener, M., Zetsche, B., Skryabin, B. V., Baba, H. A., Yanagisawa, M., and Kuhn, M. J Clin Invest More
Pressure-independent cardiac hypertrophy in mice with cardiomyocyte-restricted inactivation of the atrial natriuretic peptide receptor guanylyl cyclase-A 2003 Holtwick, R., Van Eickels, M., Skryabin, B. V., Baba, H. A., Bubikat, A., Begrow, F., Schneider, M. D., Garbers, D. L., and Kuhn, M. J Clin Invest More
Smooth muscle - selective deletion of guanylyl cyclase-A prevents the acute but not chronic effects of ANP on blood pressure 2002 Holtwick, R., Gotthardt, M., Skryabin, B., Steinmetz, M., Potthast, R., Zetsche, B., Hammer, B., Herz, J., and Kuhn, M. Proc Natl Acad Sci USA More