Purchase this article with an account.
J. I. Clark, S. Haouck, J. Ghosh; Dynamic Regulation of Microtubule Assembly by the Shsp and Molecular Chaperone, Human B Crystallin. Invest. Ophthalmol. Vis. Sci. 2008;49(13):3800. doi: https://doi.org/.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
To evaluate the function of interactive sequences in human αB crystallin on dynamic subunit exchange and microtubule assembly.
Five interactive sequences in αB crystallin were synthesized as individual peptides for experimental studies on the assembly of microtubules in vitro using a fluorescence DAPI assay (Ghosh et al. 2007 PloS one 6:3498). Mutations at corresponding positions in full-length αB crystallin were used experimentally to confirm the importance of these interactive sequences in the αB crystallin:tubulin interaction.
αB crystallin inhibited microtubule assembly at molar ratios >2:1, αB crystallin:tubulin and promoted microtubule assembly at molar ratios between 1:4 and 2:1. The plot of assembly versus molar ratio was parabolic with a maximum at approximately 0.5:1 αB:tubulin. The synthetic peptide corresponding to the αB crystallin interactive sequence, 113FISREFHR120, in the β3 strand of the α-crystallin core domain partially inhibited microtubule assembly. In contrast, the peptides 131LTITSSLSSDGV142 in the β8 strand and loop region of the core domain and 156ERTIPITRE164 in the C-terminal extension promoted microtubule assembly. Mutations in these exposed interactive sequences in full-length αB crystallin altered interactions with microtubules.
This PDF is available to Subscribers Only