Sticky salts: overbinding of monovalent cations to phosphorylations in all-atom forcefields

Phosphorylation is a major post-translational modification, which is involved in the regulation of the dynamics and function of Intrinsically Disordered Proteins (IDPs). We recently characterized a phenomenon, which we termed n -Phosphate collaborations ( n P-collabs), where bulk cations form stable bridges between several phosphoresidues in all-atom molecular dynamic simulations. n P-collabs were found to be sensitive to the combination of forcefields and cation types. Here, we attempt to assess the physical relevance of these n P-collabs by evaluating the strength of the cation/phosphate interaction through osmotic coefficient ( φ) calculations on the model 2Na ⁺ HPO ₄ ^2- and 2K ⁺ HPO ₄ ^2- salts, using different classical forcefields for phosphorylations. All forcefields were found to overestimate the strength of the interaction to various degrees. We thus designed new parameters for CHARMM36m and AmberFF99SB-ILDN using the Electronic Continuum Correction (ECC) approach, which provide remarkable agreement for φ values for both cation types and over a range of concentrations. We provide a preliminary test of these ECC parameters for phosphorylations by simulating the sevenfold-phosphorylated rhodopsin peptide 7PP and comparing secondary chemical shifts to experimental data. Conformational ensembles resulting from the ECC-derived phosphorylated forcefields display both qualitative and quantitative improvements with regards to full-charge forcefields. We thus conclude that long-lasting n P-collabs are artifacts for classical forcefields born from the lack of explicit polarization, and propose a possible computational strategy for the extensive parameterization of phosphorylations. The presence of long-lived n P-collabs in simulations produced using classical forcefields is therefore a serious concern for the accurate modelling of multiphosphorylated peptides and IDPs, which are at the center of research questions regarding neurodegenerative diseases such as Alzheimer's or Parkinson's.