Glukoserebrosidaz (GBA) genindeki kodlanan tek nükleotid polimorfizmlerinin (SNP) in siliko yöntemler ile analiz, ve hucrenin biyolojik fonksiyonları üzerindeki etkisi

Gaucher disease (GD, ORPHA355) is a rare, autosomal recessive genetic disorder. It is caused by an insufficiency of the lysosomal enzyme, glucocerebrosidase (GCase), due to the severity of GBA1 gene's mutations. Such conditions lead to GD through massive accumulation of GCase substrate, glucosylceramide, in the lysosomes. Accordingly, this research analyzed the most frequent Gaucher Disease-linked single nucleotide polymorphisms (SNPs) on the GBA1 gene by applying various bioinformatics algorithms. We have classified and characterized the L296V mutation in the GD-linked deleterious SNPs spectrum. We also showed an increase in in-vitro enzymatic activity after lysosomal reacidification and evaluated the mutant GCases' reactions to constant pH experiments. Our results showed that L296V, N370S, L444P, and D409H variants are harmful mutations with different levels of hydrolysis disturbance, tertiary structure unbalance, activation interruption, and transportation of the GCase protein as well as enzymatic efficiency reduction. Yet, we have seen lysosomes' reacidification by over-expressing hydrogen pumps (V-ATPases), which reverted the decreased stability and increased enzymatic activity. These findings may suggest a research background for the therapeutic applications of Gaucher Disease through lysosomal re-acidification, and can be extended for future studies.