Single-molecule Force Spectroscopy Predicts a Misfolded, Domain-swapped Conformation in Human γD-Crystallin Protein

Overview

Journal J Biol Chem

Specialty Biochemistry

Date 2015 Dec 26

PMID 26703476

Citations 20

Authors

Sergi Garcia-Manyes

David Giganti

Carmen L Badilla

Ainhoa Lezamiz

Judit Perales-Calvo

Amy E M Beedle

Julio M Fernandez

Affiliations

Soon will be listed here.

Abstract

Cataract is a protein misfolding disease where the size of the aggregate is directly related to the severity of the disorder. However, the molecular mechanisms that trigger the onset of aggregation remain unknown. Here we use a combination of protein engineering techniques and single-molecule force spectroscopy using atomic force microscopy to study the individual unfolding pathways of the human γD-crystallin, a multidomain protein that must remain correctly folded during the entire lifetime to guarantee lens transparency. When stretching individual polyproteins containing two neighboring HγD-crystallin monomers, we captured an anomalous misfolded conformation in which the β1 and β2 strands of the N terminus domain of two adjacent monomers swap. This experimentally elusive domain-swapped conformation is likely to be responsible for the increase in molecular aggregation that we measure in vitro. Our results demonstrate the power of force spectroscopy at capturing rare misfolded conformations with potential implications for the understanding of the molecular onset of protein aggregation.

Citing Articles

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