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Title: Properties and interactions of type III intermediate filaments with CRYAB
Author: Elliott, Jayne Louise
ISNI:       0000 0004 2735 0254
Awarding Body: Durham University
Current Institution: Durham University
Date of Award: 2013
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Glial fibrillary acidic protein is a type three intermediate filament found within astrocytes in the central nervous system and mutations have been found as the cause of Alexander disease, resulting in protein aggregates of Rosenthal fibers with αBcrystallin. Here two glial fibrillary acidic protein mutants of R79C and R239H, located in the LNDR and rod 2A domain respectively, were assembled in vitro and their morphology, assembly competence and interactions with αB-crystallin were assessed. Both mutants were unable to form usual filament lengths but instead were similar to unit length filaments with R239H forming aggregates due to such high protein interactions and the R79C mutant having much lower assembly competent protein interactions. R239H had a much greater affinity for αB-crystallin, likely a reflection that it has one of the most severe phenotypes. An absence of divalent cations equally affected GFAP assembly resulting in compromised compaction and increased filament-filament interactions. The R120G αB-crystallin mutant results in cardiomyopathy and cataract due to aberrant interactions with desmin intermediate filaments, due to an increased oligomer size and therefore these interactions were studied and compared to wild-type interactions. Temperature and pH also alter the oligomer size of αB-crystallin and were therefore investigated with αB-crystallin-type III intermediate filament interactions. It was found that ischemic conditions and increased temperatures promote their association, demonstrated by increased co-pelleting under high speed sedimentations. There was a preference for binding to desmin filaments thus showing how desmin-wild-type αB-crystallin interactions are important for homeostasis in muscle. Passive microrheology was used to complement this and investigate how αB-crystallin may be modulating desmin filaments under equilibrium. From these experiments wild-type αB-crystallin reduced the frequency-dependent passive viscosity η(f) and the G’, whereas the cardiomyopathy- and cataract- causing R120G mutant increased the η(f).
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available