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1 October 2001 The lack of chaperonelike activity of Caenorhabditis elegans Hsp12.2 cannot be restored by domain swapping with human αB-crystallin
Bas P. A. Kokke, Wilbert C. Boelens, Wilfried W. de Jong
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Abstract

The small heat shock proteins Hsp12.2 and αB-crystallin differ in that the former occurs as tetramers, without chaperonelike activity, whereas the latter forms multimers and is a good chaperone. To investigate whether the lack of chaperone activity of Hsp12.2 is primarily due to its tetrameric structure or rather to intrinsic sequence features, we engineered chimeric proteins by swapping the N-terminal, C-terminal, and tail regions of Hsp12.2 and αB-crystallin, designated as n-c-t and N-C-T, respectively. Three of the chimeric sHsps, namely N-c-T, n-c-T, and N-C-t, showed nativelike secondary and quaternary structures as measured by circular dichroism and gel permeation chromatography. Combining the conserved α-crystallin domain of Hsp12.2 with the N-terminal and tail regions of αB-crystallin (N-c-T) resulted in multimeric complexes, but did not restore chaperonelike activity. Replacing the tail region of Hsp12.2 with that of αB-crystallin (n-c-T) did not alter the tetrameric structure and lack of chaperone activity. Similarly, providing αB-crystallin with the tail of Hsp12.2 (N-C-t) did not substantially influence the multimeric complex size, but it reduced the chaperoning ability, especially for small substrates. These results suggest that the conserved α-crystallin domain of Hsp12.2 is intrinsically unsuitable to confer chaperonelike activity and confirms that the tail region in αB-crystallin modulates chaperonelike capacity in a substrate-dependent manner.

Bas P. A. Kokke, Wilbert C. Boelens, and Wilfried W. de Jong "The lack of chaperonelike activity of Caenorhabditis elegans Hsp12.2 cannot be restored by domain swapping with human αB-crystallin," Cell Stress & Chaperones 6(4), 360-367, (1 October 2001). https://doi.org/10.1379/1466-1268(2001)006<0360:TLOCAO>2.0.CO;2
Received: 8 March 2001; Accepted: 1 May 2001; Published: 1 October 2001
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