pH Induced Conformational Transitions in the Transforming Growth Factor [beta]-Induced Protein (TGF[beta]Ip) Associated Corneal Dystrophy Mutants

Most stromal corneal dystrophies are associated with aggregation and deposition of the mutated transforming growth factor-β induced protein (TGFβIp). The 4th _FAS1 domain of TGFβIp harbors ~80% of the mutations that forms amyloidogenic and non-amyloidogenic aggregates. To understand the mechanism of...

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Bibliographic Details
Published in:Scientific reports 2016-03, Vol.6, p.23836
Main Authors: Murugan, Elavazhagan, Venkatraman, Anandalakshmi, Lei, Zhou, Mouvet, Victoria, Rui Yi Lim, Rayne, Muruganantham, Nandhakumar, Goh, Eunice, Swee Lim Peh, Gary, Beuerman, Roger W, Chaurasia, Shyam S, Rajamani, Lakshminarayanan, Mehta, Jodhbir S
Format: Article
Language:English
Subjects:
Amyloidogenesis
Cornea
Corneal dystrophy
Cytotoxicity
Dystrophy
Electron microscopy
Growth factors
Light scattering
Mutants
Mutation
pH effects
Temperature effects
Thermal denaturation
Transforming growth factor-b
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Summary:Most stromal corneal dystrophies are associated with aggregation and deposition of the mutated transforming growth factor-β induced protein (TGFβIp). The 4th _FAS1 domain of TGFβIp harbors ~80% of the mutations that forms amyloidogenic and non-amyloidogenic aggregates. To understand the mechanism of aggregation and the differences between the amyloidogenic and non-amyloidogenic phenotypes, we expressed the 4th _FAS1 domains of TGFβIp carrying the mutations R555W (non-amyloidogenic) and H572R (amyloidogenic) along with the wild-type (WT). R555W was more susceptible to acidic pH compared to H572R and displayed varying chemical stabilities with decreasing pH. Thermal denaturation studies at acidic pH showed that while WT did not undergo any conformational transition, the mutants exhibited a clear pH-dependent irreversible conversion from αβ conformation to β-sheet oligomers. The β-oligomers of both mutants were stable at physiological temperature and pH. Electron microscopy and dynamic light scattering studies showed that β-oligomers of H572R were larger compared to R555W. The β-oligomers of both mutants were cytotoxic to primary human corneal stromal fibroblast (pHCSF) cells. The β-oligomers of both mutants exhibit variations in their morphologies, sizes, thermal and chemical stabilities, aggregation patterns and cytotoxicities.
ISSN:2045-2322
DOI:10.1038/srep23836