Abstract
β/γ-Crystallins are predominant structural proteins in the cytoplasm of lens fiber cells and share a similar fold composing of four Greek-key motifs divided into two domains. Numerous cataract-causing mutations have been identified in various β/γ-crystallins, but the mechanisms underlying cataract caused by most mutations remains uncharacterized. The S228P mutation in βB1-crystallin has been linked to autosomal dominant congenital nuclear cataract. Here we found that the S228P mutant was prone to aggregate and degrade in both of the human and E. coli cells. The intracellular S228P aggregates could be redissolved by lanosterol. The S228P mutation modified the refolding pathway of βB1-crystallin by affecting the formation of the dimeric intermediate but not the monomeric intermediate. Compared with native βB1-crystallin, the refolded S228P protein had less packed structures, unquenched Trp fluorophores and increased hydrophobic exposure. The refolded S228P protein was prone to aggregate at the physiological temperature and decreased the protective effect of βB1-crystallin on βA3-crystallin. Molecular dynamic simulation studies indicated that the mutation decreased the subunit binding energy and modified the distribution of surface electrostatic potentials. More importantly, the mutation separated two interacting loops in the C-terminal domain, which shielded the hydrophobic core from solvent in native βB1-crystallin. These two interacting loops are highly conserved in both of the N- and C-terminal domains of all β/γ-crystallins. We propose that these two interacting loops play an important role in the folding and structural stability of β/γ-crystallin domains by protecting the hydrophobic core from solvent access.Electronic supplementary materialThe online version of this article (doi:10.1007/s13238-016-0284-3) contains supplementary material, which is available to authorized users.
Highlights
The vertebrate lens is a delicate machine with the ability to transmit and focus visible lights on the retina
Loops in the C-terminal domain, which shielded the hydrophobic core from solvent in native βB1-crystallin. These two interacting loops are highly conserved in both of the N- and C-terminal domains of all β/γ-crystallins. We propose that these two interacting loops play an important role in the folding and structural stability of β/γ-crystallin domains by protecting the hydrophobic core from solvent access
The wild type (WT) βB1 and S228P mutant were exogenously expressed in human cell line HEK 293T and E. coli BL21 to evaluate whether the studies at the cellular and protein levels could mimic the autosomal dominant congenital cataract (ADCC) phenotype (Fig. 1)
Summary
The vertebrate lens is a delicate machine with the ability to transmit and focus visible lights on the retina. The optical functions of the lens are achieved by the three-dimensional packing of the highly differentiated lens fiber cells and the unique protein expression profile in lens cells (Benedek, 1971; Bloemendal et al, 2004). In mature lens fiber cells, all organelles have been degraded via an apoptic-like pathway to avoid light scattering (Bassnett, 2009). Once the lens proteins precipitate into large aggregates, the transmission of visible lights will be scattered, which will lead to cataract (Benedek, 1997). A typical protein aggregation disease, is one of the leading causes of human blindness worldwide. Similar to the other protein aggregation diseases, there are no clinical drugs available for cataract prevention and treatment.
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
