Back to Search
Start Over
Folding, Assembly, and Aggregation of Recombinant Murine Amelogenins with T21I and P41T Point Mutations
- Publication Year :
- 2011
- Publisher :
- S. Karger AG, 2011.
-
Abstract
- Two point mutations (T21I and P40T) within amelogenin have been identified from human DNA sequences in 2 instances of amelogenesis imperfecta. We studied the folding and self-assembly of recombinant amelogenin (rM180) compared to the T21I and P40T mutants analogs. At pH 5.8 and 25°C, rM180 and the P41T mutant existed as monomers, whereas the T21I mutant formed small oligomers. At pH 8 and 25°C, all of the amelogenin samples formed nanospheres with hydrodynamic radii (RH) of around 15–16 nm. Upon heating to 37°C, particles of P41T increased in size (RH = 18 nm). During thermal denaturation at pH 5.8, both of the mutant proteins refolded more slowly than the wild-type (WT) rM180. Variable temperature tryptophan fluorescence and dynamic light scattering studies showed that the WT transformed to a partially folded conformation upon heating and remained stable. Thermal denaturation and refolding studies indicated that the mutants were less stable and exhibit a greater ability to prematurely aggregate compared to the WT. Our data suggest that in the case of P41T, alterations in the self-assembly of amelogenin are a consequence of destabilization of the secondary structure, while in the case of T21I they are a consequence of change in the overall hydrophobicity at the N-terminal region. We propose that alterations in the assembly (i.e. premature aggregation) of mutant amelogenins may have a profound effect on intra- and extracellular processes such as amelogenin secretion, proteolysis, and its interactions with nonamelogenins as well as with the forming mineral.
- Subjects :
- Paper
Circular dichroism
Protein Folding
Histology
Mutant
Protein aggregation
Fluorescence
law.invention
Mice
stomatognathic system
law
medicine
Animals
Humans
Point Mutation
Amelogenesis imperfecta
Protein Structure, Quaternary
Protein secondary structure
Amelogenin
Chemistry
Circular Dichroism
Temperature
Hydrogen-Ion Concentration
medicine.disease
Recombinant Proteins
Biochemistry
Recombinant DNA
Protein folding
Mutant Proteins
Anatomy
Subjects
Details
- Language :
- English
- Database :
- OpenAIRE
- Accession number :
- edsair.doi.dedup.....7ec28a4d1e45f17d41a011473fab3d9d