A number of fully functional proteins have been identified to exist in a partially or fully disordered state. These intrinsically disordered proteins (IDP) are recognized as an important sequence class that fulfill many roles. A number of biomineral-associated proteins, particularly those which possess polyelectrolyte domains, represent potential members of the IDP class. This report describes a bioinformatics study of a ten member polyanionic sequence biomineralization protein family, Asprich, and the experimental characterization of the conserved N- and C-terminal regions found within seven members of this family. Using protein disorder prediction algorithms (DPROT, PONDR, GLOBPLOT), we confirm that all ten Asprich protein sequences are disordered, and that two polyelectrolyte domains within each protein contribute to the disorder scoring. Using synthetic peptides which model the conserved N- (F1,48 AA) and C-terminal (F2, 42 AA) domains, we determine that both domains are globally disordered and remain so in the presence of Ca(II). However, F1 and F2 possess differing proportions of extended beta strand relative to random coil structure and sequence spacing of Asp, Glu residues. As a result, the F2 sequence possesses a higher anionic surface charge density, solvent accessibility, and greater degree of local conformational response to Ca(II). These differences may explain why F1 and F2 differ with regard to step growth kinetics, mineral modulation, and metal ion complexation, and possibly distinguish the molecular role(s) that each domain conveys to the Asprich protein family. Structural and surface charge density features may also control the function of Asp, Glu polyelectrolyte domains within other IDP proteins.
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