May 2003
Volume 44, Issue 13
ARVO Annual Meeting Abstract  |   May 2003
Identification of Functional Domains in the -sHSP, Human B crystallin, using Protein Pin Arrays and Homology Modeling
Author Affiliations & Notes
  • J.G. Ghosh
    Biological Structure and Biomolecular Structure & Design, University of Washington, Seattle, WA, United States
  • J.I. Clark
    Biological Structure and Ophthalmology, University of Washington, Seattle, WA, United States
  • Footnotes
    Commercial Relationships  J.G. Ghosh, None; J.I. Clark, None.
  • Footnotes
    Support  EY04542
Investigative Ophthalmology & Visual Science May 2003, Vol.44, 2360. doi:
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      J.G. Ghosh, J.I. Clark; Identification of Functional Domains in the -sHSP, Human B crystallin, using Protein Pin Arrays and Homology Modeling . Invest. Ophthalmol. Vis. Sci. 2003;44(13):2360.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract: : Purpose: The molecular chaperone and small heat shock protein αB crystallin is involved in protein aggregation diseases including cataract and desmin-related myopathy (DRM). αB crystallin assembles into high molecular weight complexes having three important functional domains for subunit assembly, target protein binding and filament binding. Methods: To identify these functional domains, mimotope based protein pin arrays were used to identify sequences of human αB crystallin involved in assembly of dimers and higher order oligomers. The protein arrays consisted of overlapping peptides spanning the entire sequence of human αB crystallin and were synthesized in a multi-well ELISA plate format. Homology models for human αB crystallin based on the 2.9Å crystal structure of the wheat HSP 16.9 were constructed (refer figure). Results: The peptides 41STSLSPFYLRPPSFLRAP58, 73DRFSVNLDVK82 and 131LTITSSLSSDGV142 belonging to the N-terminus, core "α-crystallin" domain and the C-terminus extension respectively bind strongly to human αB crystallin and human αA crystallin and appear to be involved in subunit assembly. None of the binding sequences belonged to the loop region (residues 100-121) that has been identified as one of the dimer interfaces in the crystal structures of the wheat HSP 16.9. and the Mj HSP16.5. All sequences identified as functional domains and mapped onto homology models were solvent exposed and had variable secondary structures ranging from ß-strands to random coils and short α-helices.  

Keywords: cataract • protein structure/function • protein purification and characterization 

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