Multi-Tool Computational Analysis of Mutational Tolerance at Critical Human BEST1 Bestrophin-1 Residues..."

Abstract Background: Bestrophinopathies, including Best Vitelliform Macular Dystrophy (BVMD), are caused by mutations in the BEST1 gene. Despite its clinical importance, the mutational tolerance of many critical residues remain poorly understood, particularly in South Asian populations where clinical data is often underrepresented. Methods: This study utilizes an integrated in silico framework to evaluate the impact of amino acid substitutions on the Bestrophin-1 protein (PDB: 6N28). We employed a six-component computational pipeline—including PolyPhen-2 for functional impact, and DynaMut2 and mCSM for protein stability—to analyze high-risk variants. Structural models were validated using SAVES v6.1 (ERRAT and Ramachandran plots). Results: Our analysis achieved 100% concordance with known human clinical variants (e.g., W93C, P297S) obtained from ClinVar and HGMD. We identified specific mutational hotspots at critical residues where amino acid substitutions significantly disrupt the protein's three-dimensional discordance and stability. The structural validation scores (ERRAT: 88.162) confirm the high reliability of the predicted deleterious effects.