Identification of the Most Impactful Asparagine Residues for γS-Crystallin Aggregation by Deamidation.

Koichi Kato, Tomoki Nakayoshi, Yuki Kitamura, Eiji Kurimoto, Akifumi Oda, Yoshinobu Ishikawa

Biochemistry 2023 Jun 06

filter terms:

Crystallin aggregation in the eye lens is involved in the pathogenesis of cataracts. The aggregation is considered to be promoted by non-enzymatic post-translational modifications, such as the deamidation and stereoinversion of amino acid residues. Although in a previous study, the deamidated asparagine residues were detected in γS-crystallin in vivo, it is unclear which deamidated residues have the most impact on the aggregation under physiological conditions. In this study, we investigated the deamidation impacts of all Asn residues in γS-crystallin for the structural and aggregation properties utilizing deamidation mimetic mutants (N14D, N37D, N53D, N76D, and N143D). The structural impacts were investigated using circular dichroism analysis and molecular dynamics simulations, and the aggregation properties were analyzed by gel filtration chromatography and spectrophotometric methods. No significant structural impacts of all mutations were detected. However, the N37D mutation decreased thermal stability and changed some intermolecular hydrogen-bond formations. Aggregation analysis indicated that the superiority of the aggregation rate in each mutant varied with temperature. Deamidation at any Asn residues promoted γS-crystallin aggregation, and the deamidation at Asn37, Asn53, and Asn76 were suggested to be the most impactful in the formation of insoluble aggregations.

Citation

Koichi Kato, Tomoki Nakayoshi, Yuki Kitamura, Eiji Kurimoto, Akifumi Oda, Yoshinobu Ishikawa. Identification of the Most Impactful Asparagine Residues for γS-Crystallin Aggregation by Deamidation. Biochemistry. 2023 Jun 06;62(11):1679-1688