DETECTION OF MONODISPERSE AGGREGATES OF A RECOMBINANT AMELOGENIN BY DYNAMIC LIGHT-SCATTERING

被引:120
作者
MORADIANOLDAK, J
SIMMER, JP
LAU, EC
SARTE, PE
SLAVKIN, HC
FINCHAM, AG
机构
[1] UNIV SO CALIF,SCH DENT,CTR CRANIOFACIAL MOLEC BIOL,LOS ANGELES,CA 90033
[2] UNIV TEXAS,HLTH SCI CTR,SCH DENT,SAN ANTONIO,TX 78284
关键词
D O I
10.1002/bip.360341006
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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. (C) 1994 John Wiley and Sons, Inc.
引用
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页码:1339 / 1347
页数:9
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