Site-Specific Phosphorylation of PSD-95 PDZ Domains Reveals Fine-Tuned Regulation of Protein-Protein Interactions
Research output: Contribution to journal › Journal article › Research › peer-review
The postsynaptic density protein of 95 kDa (PSD-95) is a key scaffolding protein that controls signaling at synapses in the brain through interactions of its PDZ domains with the C-termini of receptors, ion channels, and enzymes. PSD-95 is highly regulated by phosphorylation. To explore the effect of phosphorylation on PSD-95, we used semisynthetic strategies to introduce phosphorylated amino acids at four positions within the PDZ domains and examined the effects on interactions with a large set of binding partners. We observed complex effects on affinity. Most notably, phosphorylation at Y397 induced a significant increase in affinity for stargazin, as confirmed by NMR and single molecule FRET. Additionally, we compared the effects of phosphorylation to phosphomimetic mutations, which revealed that phosphomimetics are ineffective substitutes for tyrosine phosphorylation. Our strategy to generate site-specifically phosphorylated PDZ domains provides a detailed understanding of the role of phosphorylation in the regulation of PSD-95 interactions.
|Journal||ACS chemical biology|
|Number of pages||11|
|Publication status||Published - 15 Sep 2017|
- Amino Acid Sequence, Calcium Channels, Disks Large Homolog 4 Protein, Humans, Intracellular Signaling Peptides and Proteins, Membrane Proteins, Models, Molecular, PDZ Domains, Phosphorylation, Protein Folding, Protein Interaction Maps, Protein Stability, Journal Article