Computational Tools Unravel Putative Sterol Binding Sites in the Lysosomal NPC1 Protein

• Published in: Journal of chemical information and modeling 2019-05, Vol.59 (5), p.2432-2441
• Main Author: Elghobashi-Meinhardt, Nadia
• Format: Article
• Language: English
• Subjects: Amino acids; Binding sites; Cholesterol; Computer simulation; Critical components; Domains; Lipids; Molecular docking; Molecular dynamics; Mutation; Proteins; Protonation; Software; Sterols
• ISSN: 1549-9596; 1549-960X
• DOI: 10.1021/acs.jcim.9b00186

Two proteins have been linked as the critical components in the molecular mechanisms involved in the Niemann Pick type C (NPC) disease: NPC1, a 140 kDa polytopic membrane-bound protein, and the smaller (132 residues), water-soluble NPC2 protein. NPC1 is believed to act in tandem with NPC2, transferring cholesterol and other sterols out of the LE/Lys compartments. Mutations in either NPC1 or NPC2 can lead to an accumulation of cholesterol and lipids in the LE/Lys, the primary phenotype of the NPC disease, but approximately 95% of identified disease-causing mutations have been mapped to the membrane-bound NPC1 protein. Here, we investigate the full length, membrane-bound NPC1 protein computationally using a combination of molecular modeling, docking, and molecular dynamics (MD) simulations. An analysis of titratable amino acid side chains, several buried in protein pockets, reveals several nonstandard protonation states for the low-pH scenario (pH 5) that is realized in the lysosome. Together with the location of these buried amino acids, docking studies have identified putative lipid binding domains that are in close proximity to amino acids that, when mutated, are connected to NPC1 loss-of-function. Using energy analyses and MD simulations, we analyze these domains as potential cholesterol binding sites and propose the possibility of multiple sterol binding pockets enabling the intramolecular transport of sterol molecules to the transmembrane domain.