In vitro effects of recombinant otoconin 90 upon calcite crystal growth. Significance of tertiary structure

• Published in: Hearing research 2010-09, Vol.268 (1), p.172-183
• Main Authors: Lu, Wenfu, Zhou, Dan, Freeman, John J., Thalmann, Isolde, Ornitz, David M., Thalmann, Ruediger
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Animals; Biological and medical sciences; Calcium Carbonate - chemistry; Calcium Carbonate - metabolism; Cell Line; Circular Dichroism; Crystallization; Drug toxicity and drugs side effects treatment; Extracellular Matrix Proteins - chemistry; Extracellular Matrix Proteins - genetics; Extracellular Matrix Proteins - metabolism; Humans; Kinetics; Light; Medical sciences; Mice; Models, Molecular; Molecular Sequence Data; Otolithic Membrane - chemistry; Otolithic Membrane - metabolism; Pharmacology. Drug treatments; Protein Folding; Protein Structure, Tertiary; Recombinant Proteins - chemistry; Scattering, Radiation; Spectrum Analysis, Raman; Structural Homology, Protein; Structure-Activity Relationship; Toxicity: respiratory system, ent, stomatology; Transfection
• ISSN: 0378-5955; 1878-5891
• DOI: 10.1016/j.heares.2010.05.019

Otoconia are biomineral particles of microscopic size essential for perception of gravity and maintenance of balance. Millions of older Americans are affected in their mobility, quality of life and in their health by progressive demineralization of otoconia. Currently, no effective means to prevent or counteract this process are available. Because of prohibitive anatomical and biological constraints, otoconial research is lagging far behind other systems such as bone and teeth. We have overcome these obstacles by generating otoconial matrix proteins by recombinant techniques. In the present study, we evaluated the effects of recombinant Otoconin 90 (OC90), the principal soluble matrix protein upon calcite crystal growth patterns in vitro. Our findings highlight multiple effects, including facilitation of nucleation, and inhibition of crystal growth in a concentration-dependent manner. Moreover, OC90 induces morphologic changes characteristic of native otoconia. OC90 is considerably less acidic than the prototypical invertebrate CaCO 3 –associated protein, but is nevertheless an effective modulator of calcite crystal growth. Based on homology modeling of the sPLA2-like domains of OC90, we propose that the lower density of acidic residues of the primary sequence is compensated by formation of major anionic surface clusters upon folding into tertiary conformation.