Next Article in Journal
Photodynamic and Cold Atmospheric Plasma Combination Therapy Using Polymeric Nanoparticles for the Synergistic Treatment of Cervical Cancer
Next Article in Special Issue
Update on the Roles of Rice MAPK Cascades
Previous Article in Journal
Expression of Flavodiiron Proteins Flv2-Flv4 in Chloroplasts of Arabidopsis and Tobacco Plants Provides Multiple Stress Tolerance
Previous Article in Special Issue
Differential Role of Threonine and Tyrosine Phosphorylation in the Activation and Activity of the Yeast MAPK Slt2
Article

Identification of Novel Substrates for cGMP Dependent Protein Kinase (PKG) through Kinase Activity Profiling to Understand Its Putative Role in Inherited Retinal Degeneration

1
Division of Toxicology, Wageningen University and Research, Stippeneng 4, 6708 WE Wageningen, The Netherlands
2
PamGene International B.V., P.O. Box 1345, 5200 BJ ‘s-Hertogenbosch, The Netherlands
3
Department of Life Sciences, University of Modena and Reggio Emilia, via Campi, 287, 41125 Modena, Italy
4
Center for Neuroscience and Neurotechnology, University of Modena and Reggio Emilia, via Campi, 287, 41125 Modena, Italy
*
Author to whom correspondence should be addressed.
Academic Editor: Atsushi Matsuzawa
Int. J. Mol. Sci. 2021, 22(3), 1180; https://doi.org/10.3390/ijms22031180
Received: 28 December 2020 / Revised: 19 January 2021 / Accepted: 20 January 2021 / Published: 25 January 2021
(This article belongs to the Special Issue Kinase Signal Transduction 2.0)
Inherited retinal degenerative diseases (IRDs), which ultimately lead to photoreceptor cell death, are characterized by high genetic heterogeneity. Many IRD-associated genetic defects affect 3′,5′-cyclic guanosine monophosphate (cGMP) levels. cGMP-dependent protein kinases (PKGI and PKGII) have emerged as novel targets, and their inhibition has shown functional protection in IRDs. The development of such novel neuroprotective compounds warrants a better understanding of the pathways downstream of PKGs that lead to photoreceptor degeneration. Here, we used human recombinant PKGs in combination with PKG activity modulators (cGMP, 3′,5′-cyclic adenosine monophosphate (cAMP), PKG activator, and PKG inhibitors) on a multiplex peptide microarray to identify substrates for PKGI and PKGII. In addition, we applied this technology in combination with PKG modulators to monitor kinase activity in a complex cell system, i.e. the retinal cell line 661W, which is used as a model system for IRDs. The high-throughput method allowed quick identification of bona fide substrates for PKGI and PKGII. The response to PKG modulators helped us to identify, in addition to ten known substrates, about 50 novel substrates for PKGI and/or PKGII which are either specific for one enzyme or common to both. Interestingly, both PKGs are able to phosphorylate the regulatory subunit of PKA, whereas only PKGII can phosphorylate the catalytic subunit of PKA. In 661W cells, the results suggest that PKG activators cause minor activation of PKG, but a prominent increase in the activity of cAMP-dependent protein kinase (PKA). However, the literature suggests an important role for PKG in IRDs. This conflicting information could be reconciled by cross-talk between PKG and PKA in the retinal cells. This must be explored further to elucidate the role of PKGs in IRDs. View Full-Text
Keywords: PKG; PKA; cGMP; cAMP; 661W; retinal degeneration; substrate identification; peptide microarray PKG; PKA; cGMP; cAMP; 661W; retinal degeneration; substrate identification; peptide microarray
Show Figures

Figure 1

MDPI and ACS Style

Roy, A.; Groten, J.; Marigo, V.; Tomar, T.; Hilhorst, R. Identification of Novel Substrates for cGMP Dependent Protein Kinase (PKG) through Kinase Activity Profiling to Understand Its Putative Role in Inherited Retinal Degeneration. Int. J. Mol. Sci. 2021, 22, 1180. https://doi.org/10.3390/ijms22031180

AMA Style

Roy A, Groten J, Marigo V, Tomar T, Hilhorst R. Identification of Novel Substrates for cGMP Dependent Protein Kinase (PKG) through Kinase Activity Profiling to Understand Its Putative Role in Inherited Retinal Degeneration. International Journal of Molecular Sciences. 2021; 22(3):1180. https://doi.org/10.3390/ijms22031180

Chicago/Turabian Style

Roy, Akanksha, John Groten, Valeria Marigo, Tushar Tomar, and Riet Hilhorst. 2021. "Identification of Novel Substrates for cGMP Dependent Protein Kinase (PKG) through Kinase Activity Profiling to Understand Its Putative Role in Inherited Retinal Degeneration" International Journal of Molecular Sciences 22, no. 3: 1180. https://doi.org/10.3390/ijms22031180

Find Other Styles
Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Article Access Map by Country/Region

1
Back to TopTop