alphaB-crystallin - the core and the Oligomer : A Structural Investigation
EMSL Project ID
10297
Abstract
alphaB-crystallin (alphaB) 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. Wild type (w.t.) alphaB is assembled from 175 aa, 20 kDa subunits to form a polydisperse oligomer with the molecular weight in the 650 ± 150 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 w.t. alphaB called alphaB57. alphaB57 undergoes a pH-dependent structural/conformational change. We would like to acquire NOESY acquire at a higher field strength, viz., 800 MHz to enable us to calculate high-resolution structures at different pH values. We would like to use 900 MHz NMR to implement experiments suitable for supramolecular systems, e.g., CRINEPT-TROSY and CRIPT-TROSY. We would then acquire these spectra on w.t. alphaB to obtain some backbone amide chemical shift data.
Project Details
Project type
Capability Research
Start Date
2004-11-10
End Date
2005-10-03
Status
Closed
Released Data Link
Team
Principal Investigator
Team Members