Synergistic Biomineralization Phenomena Created by a Combinatorial Nacre Protein Model System

Eric P. Chang, Teresa Roncal-Herrero, Tamara Morgan, Katherine E. Dunn, Ashit Rao, Jennie A.M.R. Kunitake, Susan Lui, Matthew Bilton, Lara A. Estroff, Roland Kröger, Steven Johnson, Helmut Cölfen, John Spencer Evans

    Research output: Contribution to journalArticlepeer-review


    In the nacre or aragonite layer of the mollusk shell, proteomes that regulate both the early stages of nucleation and nano-to-mesoscale assembly of nacre tablets from mineral nanoparticle precursors exist. Several approaches have been developed to understand protein-associated mechanisms of nacre formation, yet we still lack insight into how protein ensembles or proteomes manage nucleation and crystal growth. To provide additional insights, we have created a proportionally defined combinatorial model consisting of two nacre-associated proteins, C-RING AP7 (shell nacre, Haliotis rufescens) and pseudo-EF hand PFMG1 (oyster pearl nacre, Pinctada fucata), whose individual in vitro mineralization functionalities are well-documented and distinct from one another. Using scanning electron microscopy, flow cell scanning transmission electron microscopy, atomic force microscopy, Ca(II) potentiometric titrations, and quartz crystal microbalance with dissipation monitoring quantitative analyses, we find that both nacre proteins are functionally active within the same mineralization environments and, at 1:1 molar ratios, synergistically create calcium carbonate mesoscale structures with ordered intracrystalline nanoporosities, extensively prolong nucleation times, and introduce an additional nucleation event. Further, these two proteins jointly create nanoscale protein aggregates or phases that under mineralization conditions further assemble into protein-mineral polymer-induced liquid precursor-like phases with enhanced ACC stabilization capabilities, and there is evidence of intermolecular interactions between AP7 and PFMG1 under these conditions. Thus, a combinatorial model system consisting of more than one defined biomineralization protein dramatically changes the outcome of the in vitro biomineralization process.

    Original languageEnglish (US)
    Pages (from-to)2401-2410
    Number of pages10
    Issue number16
    StatePublished - May 10 2016

    ASJC Scopus subject areas

    • Biochemistry


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