The leucine rich amelogenin protein (LRAP) adsorbs as monomers or dimers onto surfaces.
Amelogenin is believed to be involved in controlling the formation of the highly anisotropic and ordered hydroxyapatite crystallites that form enamel. The adsorption behavior of amelogenin proteins onto substrates is very important because protein-surface interactions are critical to its function. We have previously used LRAP, a splice variant of amelogenin, as a model protein for the full-length amelogenin in solid-state NMR and neutron reflectivity studies at interfaces. In this work, we examined the adsorption behavior of LRAP in greater detail using model self-assembled monolayers containing COOH, CH(3), and NH(2) end groups as substrates. Dynamic light scattering (DLS) experiments indicated that LRAP in phosphate buffered saline and solutions containing low concentrations of calcium and phosphate consisted of aggregates of nanospheres. Null ellipsometry and atomic force microscopy (AFM) were used to study protein adsorption amounts and quaternary structures on the surfaces. Relatively high amounts of adsorption occurred onto the CH(3) and NH(2) surfaces from both buffer solutions. Adsorption was also promoted onto COOH surfaces only when calcium was present in the solutions suggesting an interaction that involves calcium bridging with the negatively charged C-terminus. The ellipsometry and AFM studies revealed that LRAP adsorbed onto the surfaces as small subnanosphere-sized structures such as monomers or dimers. We propose that the monomers/dimers were present in solution even though they were not detected by DLS or that they adsorbed onto the surfaces by disassembling or "shedding" from the nanospheres that are present in solution. This work reveals the importance of small subnanosphere-sized structures of LRAP at interfaces.
Pacific Northwest National Laboratory, 908 Battelle Blvd., Richland, WA 99352, USA. barbara.tarasevich@pnl.gov
Barbara J
BJ
Tarasevich
Scott
S
Lea
Wendy J
WJ
Shaw
D000327:Adsorption; D053523:Amelogenin / Q000138:chemical synthesis / Q000737:chemistry*; D000818:Animals; D051379:Mice; D018625:Microscopy, Atomic Force; D055503:Protein Multimerization; D012542:Scattering, Radiation
D053523:Amelogenin