Exploring the Mechanism of Action of Membrane Fusion Protein IncA Using Molecular Dynamics Simulations

Abstract

Infecting millions of people each year, Chlamydia trachomatis is currently the leading cause of non‐congenital blindness and pelvic inflammatory disease. Incapable of replicating on its own, this bacterium survives by entering a host cell through endocytosis and subsequently modifying the vacuole in which it is enclosed to allow for replication. The newly formed replicative compartment, referred to as an inclusion, has numerous inclusion membrane proteins (Inc proteins) that promote growth and maintenance of the developing C. trachomatis bodies. One Inc in particular, known as IncA, exponentially enhances growth of C. trachomatis by initiating the homotypic fusion of multiple inclusions within a host cell. Although the structure and significance of IncA are known, its mechanism of action remains a mystery. The present research aims to elucidate the structure‐function relationship of IncA; offering insight into fusion protein functionality and potential targets for drug development. Analysis of IncA was accomplished by running Constant pH Molecular Dynamics (CpH MD) simulations with a refined focus on aspartic acids, glutamic acids, and histidines. Similar to conventional MD, CpH MD creates conditions that mimic the natural environment of IncA but with the additional capability to explore protonation states of residues. By running the CpH MD system across a range of pHs, the necessary data are generated to determine dissociation constants for specific residues. Combining proton dissociation with tests examining the solvent accessibility of IncA residues, results suggested that there is a correlation between accessibility of a residue and its proton dissociation. Further analysis of residue fluctuation at varying pHs shows that there appears to be a relationship between a residue's protonation state and the degree to which that residue fluctuated throughout a simulation. These findings demonstrate that the location of titratable residues influence the activity of IncA and furthermore, that IncA's titratable residues affect inclusion membrane fusion.Support or Funding InformationNational Science Foundation, Grant No. 1560325.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.