Folding, stability and aggregation of the long-lived eye lens protein human gamma D crystallin

Human [gamma]D crystallin (H[gamma]D-Crys) is a monomeric, two domain, primarily P-sheet protein found in high concentrations in the human eye lens. H[gamma]D-Crys and other crystallins are found in insoluble protein inclusions associated with the eye disease cataract. H[gamma]D-Crys is expressed in utero and does not regenerate during life, thus necessitating high stability and solubility. Covalent damage, including glutamine deamidation, of the lens crystallins increases with age and as a result of exposure to environmental insults. Such covalent damage may cause partial-unfolding into aggregation-prone confomations that cause cataract. The in vitro stability of H[gamma]D-Crys was analyzed in the denaturant guanidine h[gamma]Drochloride at pH 7.0 and 370C. An off-pathway aggregation reaction that competed with refolding was previously reported when H[gamma]D-Crys was refolded to less than 1 M GuHC1. Equilibrium transitions of H[gamma]D-Crys were best fit to a three-state model suggesting the presence of a partially-folded intermediate that likely had a structured C-terminal domain (C-td) and unstructured N-terminal domain (N-td). Similarly, previous analyses revealed a sequential domain refolding pathway where the C-td refolds first followed by the N-td.

Department dc:contributor.department

Massachusetts Institute of Technology. Dept. of Biology.

Grantor dc:publisher

Massachusetts Institute of Technology

Year dc:date.issued

2006

Author dc:creator

• Flaugh, Shannon L

Advisor dc:contributor.advisor

• Jonathan King.

• Biology.

Statement dc:rights

• M.I.T. theses are protected by copyright. They may be viewed from this source for any purpose, but reproduction or distribution in any format is prohibited without written permission. See provided URL for inquiries about permission.

Licence dc:rights.uri

• http://dspace.mit.edu/handle/1721.1/34572
• http://dspace.mit.edu/handle/1721.1/7582

Language dc:language.iso

eng

Identifier URI

http://dspace.mit.edu/handle/1721.1/34572

OAI identifier oai:identifier

oai:dspace.mit.edu:1721.1/34572