Light-dependent phosphorylation of the Drosophila inactivation no afterpotential D (INAD) scaffolding protein at Thr170 and Ser174 by eye-specific protein kinase C

Olaf Voolstra; Philipp Spät; Claudia Oberegelsbacher; Björn Claussen; Jens Pfannstiel; Armin Huber

doi:10.1371/journal.pone.0122039

Light-dependent phosphorylation of the Drosophila inactivation no afterpotential D (INAD) scaffolding protein at Thr170 and Ser174 by eye-specific protein kinase C

PLoS One. 2015 Mar 23;10(3):e0122039. doi: 10.1371/journal.pone.0122039. eCollection 2015.

Authors

Olaf Voolstra¹, Philipp Spät¹, Claudia Oberegelsbacher¹, Björn Claussen¹, Jens Pfannstiel², Armin Huber³

Affiliations

¹ Department of Biosensorics, Institute of Physiology, Universität Hohenheim, Stuttgart, Germany.
² Mass Spectrometry Core Facility, Universität Hohenheim, Stuttgart, Germany.
³ Department of Biosensorics, Institute of Physiology, Universität Hohenheim, Stuttgart, Germany; Mass Spectrometry Core Facility, Universität Hohenheim, Stuttgart, Germany.

Abstract

Drosophila inactivation no afterpotential D (INAD) is a PDZ domain-containing scaffolding protein that tethers components of the phototransduction cascade to form a supramolecular signaling complex. Here, we report the identification of eight INAD phosphorylation sites using a mass spectrometry approach. PDZ1, PDZ2, and PDZ4 each harbor one phosphorylation site, three phosphorylation sites are located in the linker region between PDZ1 and 2, one site is located between PDZ2 and PDZ3, and one site is located in the N-terminal region. Using a phosphospecific antibody, we found that INAD phosphorylated at Thr170/Ser174 was located within the rhabdomeres of the photoreceptor cells, suggesting that INAD becomes phosphorylated in this cellular compartment. INAD phosphorylation at Thr170/Ser174 depends on light, the phototransduction cascade, and on eye-Protein kinase C that is attached to INAD via one of its PDZ domains.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Sequence
Animals
Animals, Genetically Modified
Drosophila / physiology*
Drosophila Proteins / chemistry
Drosophila Proteins / genetics
Drosophila Proteins / metabolism*
Eye Proteins / chemistry
Eye Proteins / genetics
Eye Proteins / metabolism*
Light Signal Transduction*
Light*
Molecular Sequence Data
Mutation
Phosphorylation
Protein Kinase C / chemistry
Protein Kinase C / genetics
Protein Kinase C / metabolism*

Substances

Drosophila Proteins
Eye Proteins
inaD protein, Drosophila
Protein Kinase C

Grants and funding

Funding was provided by Deutsche Forschungsgemeinschaft (VO 1741/1-1) to OV; German-Israeli-Foundation for Research and Development (G.I.F. grant I-1001-96.13) to OV, AH. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.