Structural Investigation of a Molecular Chaperone
EMSL Project ID
12990
Abstract
?B-Crystallin (?B) belongs to the family of small heat shock proteins (sHSPs) and has been shown in vitro to be a molecular chaperone. It is one of the proteins essential for maintaining eye-lens transparency and is found associated with a variety of neurodegenerative diseases, e.g., Alzheimer?s disease. ?B is assembled from 175 aa, 20 kDa subunits to form a polydisperse oligomer with the molecular weight in ~580 kDa range. We propose to tackle the structure of the oligomer in incremental steps starting with the 100-residue core-domain which is common to all sHSPs. We have cloned and expressed in E. coli, a 100-residue core domain of ?B called ?B58. ?B58 undergoes a pH-dependent structural/conformational change. We would like to acquire triple-resonance spectra at 600 MHz to assign the resonances of the two conformations. We would like to use 800 MHz to investigate the binding of ?B58 to a denatured substrate to begin to elucidate the chaperoning mechanism. We would like to use 900 MHz NMR supramolecular to acquire CRINEPT-TROSY and CRIPT-TROSY spectra on ?B to obtain some backbone amide chemical shift data.
Project Details
Project type
Capability Research
Start Date
2005-05-05
End Date
2005-10-03
Status
Closed
Released Data Link
Team
Principal Investigator
Team Members