Detection of monodisperse aggregates of a recombinant amelogenin by dynamic light scattering.
Recombinant murine amelogenins M179 and M166 were expressed in Escherichia coli and purified. The aggregation properties of these amelogenins have been investigated in aqueous solutions as well as acetonitrile-containing solutions using dynamic light scattering. Dynamic light scattering provides direct measurement of the translational diffusion coefficient and hydrodynamic radius, and of an estimate of the molecular weight. Polydispersity and statistical parameters of how to interpret the analysis are also provided. Amelogenin aggregation was examined in solutions of a range of pH, ionic strengths, and protein concentrations. It was shown that at pH 7.8-8 and ionic strength of 0.02-0.05M the M179 molecules form monodispersed aggregates with hydrodynamic radii ranging from 15 to 19 nm. Analysis of hydrodynamic radii and size distribution of M179 aggregates in acetonitrile-containing solvents compared to that in aqueous solutions indicated a primary role for hydrophobic interactions in the association process of amelogenin molecules to form aggregates. Comparison between the aggregates formed by M179 and M166, which lacks the hydrophilic carboxy-terminal 13 residue sequence of M179, suggested that the self-assembly of amelogenin molecules to form stable and monodisperse aggregates requires the presence of the hydrophilic carboxy-terminal sequence of M179.
Moradian-Oldak
J
J
Center for Craniofacial Molecular Biology, School of Dentistry, University of Southern California, Los Angeles 90033.
Simmer
J P
JP
Lau
E C
EC
Sarte
P E
PE
Slavkin
H C
HC
Fincham
A G
AG
D053523:Amelogenin; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D003746:Dental Enamel Proteins / Q000737:chemistry*; D008027:Light; D051379:Mice; D008969:Molecular Sequence Data; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry; D012542:Scattering, Radiation
D053523:Amelogenin; C507200:Amelx protein, mouse; D003746:Dental Enamel Proteins; D011994:Recombinant Proteins