Amelogenin Supramolecular Assembly in Nanospheres Defined by a Complex Helix-Coil-PPII Helix 3D-Structure

Tooth enamel, the hardest material in the human body, is formed within a self-assembled matrix consisting mostly of amelogenin proteins. Here we have determined the complete mouse amelogenin structure under physiological conditions and defined interactions between individual domains. NMR spectroscopy revealed four major amelogenin structural motifs, including an N-terminal assembly of four α-helical segments (S9-V19, T21-P33, Y39-W45, V53-Q56), an elongated random coil region interrupted by two 310 helices (∼P60-Q117), an extended proline-rich PPII-helical region (P118-L165), and a charged hydrophilic C-terminus (L165-D180). HSQC experiments demonstrated ipsilateral interactions between terminal domains of individual amelogenin molecules, i.e. N-terminal interactions with corresponding N-termini and C-terminal interactions with corresponding C-termini, while the central random coil domain did not engage in interactions. Our HSQC spectra of the full-length amelogenin central domain region completely overlapped with spectra of the monomeric Amel-M fragment, suggesting that the central amelogenin coil region did not involve in assembly, even in assembled nanospheres. This finding was confirmed by analytical ultracentrifugation experiments. We conclude that under conditions resembling those found in the developing enamel protein matrix, amelogenin molecules form complex 3D-structures with N-terminal α-helix-like segments and C-terminal PPII-helices, which self-assemble through ipsilateral interactions at the N-terminus of the molecule

Climatic and geologic controls on the piezometry of the Querença-Silves karst aquifer, Algarve (Portugal)

Karst aquifers in semi-arid regions, like Queren double dagger a-Silves (Portugal), are particularly vulnerable to climate variability. For the first time in this region, the temporal structure of a groundwater-level time series (1985-2010) was explored using the continuous wavelet transform. The investigation focused on a set of four piezometers, two at each side of the S. Marcos-Quarteira fault, to demonstrate how each of the two sectors of the aquifer respond to climate-induced patterns. Singular spectral analysis applied to an extended set of piezometers enabled identification of several quasi-periodic modes of variability, with periods of 6.5, 4.3, 3.2 and 2.6 years, which can be explained by low-frequency climate patterns. The geologic forcing accounts for similar to 15 % of the differential variability between the eastern and western sectors of the aquifer. The western sector displays spatially homogenous piezometric variations, large memory effects and low-pass filtering characteristics, which are consistent with relatively large and uniform values of water storage capacity and transmissivity properties. In this sector, the 6.5-year mode of variability accounts for similar to 70 % of the total variance of the groundwater levels. The eastern sector shows larger spatial and temporal heterogeneity, is more reactive to short-term variations, and is less influenced by the low-frequency components related to climate patterns