R120G alpha B-crystallin promotes the unfolding of reduced alpha-lactalbumin and is inherently unstable

Treweek, Teresa M., Rekas, Agata, Lindner, Robyn A., Walker, Mark J., Aquilina, J. Andrew, Robinson, Carol V., Horwitz, Joseph, Perng, Ming Der, Quinlan, Roy A. and Carver, John A. (2005) R120G alpha B-crystallin promotes the unfolding of reduced alpha-lactalbumin and is inherently unstable. FEBS Journal, 272 3: 711-724. doi:10.1111/j.1742-4658.2004.04507.x


Author Treweek, Teresa M.
Rekas, Agata
Lindner, Robyn A.
Walker, Mark J.
Aquilina, J. Andrew
Robinson, Carol V.
Horwitz, Joseph
Perng, Ming Der
Quinlan, Roy A.
Carver, John A.
Title R120G alpha B-crystallin promotes the unfolding of reduced alpha-lactalbumin and is inherently unstable
Formatted title
R120G αB-crystallin promotes the unfolding of reduced α-lactalbumin and is inherently unstable
Journal name FEBS Journal   Check publisher's open access policy
ISSN 1742-464X
1742-4658
Publication date 2005-02
Sub-type Article (original research)
DOI 10.1111/j.1742-4658.2004.04507.x
Volume 272
Issue 3
Start page 711
End page 724
Total pages 14
Place of publication Oxford, United Kingdom
Publisher Wiley-Blackwell Publishing
Language eng
Formatted abstract
α-Crystallin is the principal lens protein which, in addition to its structural role, also acts as a molecular chaperone, to prevent aggregation and precipitation of other lens proteins. One of its two subunits, αB-crystallin, is also expressed in many nonlenticular tissues, and a natural missense mutation, R120G, has been associated with cataract and desmin-related myopathy, a disorder of skeletal muscles [Vicart P, Caron A, Guicheney P, Li Z, Prevost MC, Faure A, Chateau D, Chapon F, Tome F, Dupret JM, Paulin D & Fardeau M (1998) Nat Genet20, 92–95]. In the present study, real-time 1H-NMR spectroscopy showed that the ability of R120G αB-crystallin to stabilize the partially folded, molten globule state of α-lactalbumin was significantly reduced in comparison with wild-type αB-crystallin. The mutant showed enhanced interaction with, and promoted unfolding of, reduced α-lactalbumin, but showed limited chaperone activity for other target proteins. Using NMR spectroscopy, gel electrophoresis, and MS, we observed that, unlike the wild-type protein, R120G αB-crystallin is intrinsically unstable in solution, with unfolding of the protein over time leading to aggregation and progressive truncation from the C-terminus. Light scattering, MS, and size-exclusion chromatography data indicated that R120G αB-crystallin exists as a larger oligomer than wild-type αB-crystallin, and its size increases with time. It is likely that removal of the positive charge from R120 of αB-crystallin causes partial unfolding, increased exposure of hydrophobic regions, and enhances its susceptibility to proteolysis, thus reducing its solubility and promoting its aggregation and complexation with other proteins. These characteristics may explain the involvement of R120G αB-crystallin with human disease states.
Keyword Cataract
Lens proteins
Molecular chaparone
Protein aggregation
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Non-UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: ERA 2012 Admin Only
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