Glutamate transporter homolog-based model predicts that anion-π interaction is the mechanism for the voltage-dependent response of prestin.
Prestin is the motor protein of cochlear outer hair cells. Its unique capability to perform direct, rapid, and reciprocal electromechanical conversion depends on membrane potential and interaction with intracellular anions. How prestin senses the voltage change and interacts with anions are still unknown. Our three-dimensional model of prestin using molecular dynamics simulations predicts that prestin contains eight transmembrane-spanning segments and two helical re-entry loops and that tyrosyl residues are the structural specialization of the molecule for the unique function of prestin. Using site-directed mutagenesis and electrophysiological techniques, we confirmed that residues Tyr(367), Tyr(486), Tyr(501), and Tyr(508) contribute to anion binding, interacting with intracellular anions through novel anion-π interactions. Such weak interactions, sensitive to voltage and mechanical stimulation, confer prestin with a unique capability to perform electromechanical and mechanoelectric conversions with exquisite sensitivity. This novel mechanism is completely different from all known mechanisms seen in ion channels, transporters, and motor proteins.
From the Department of Biomedical Sciences, Creighton University School of Medicine, Omaha, Nebraska 68178 slovas@creighton.edu.
Sándor
S
Lovas
From the Department of Biomedical Sciences, Creighton University School of Medicine, Omaha, Nebraska 68178 davidhe@creighton.edu.
David Z Z
DZ
He
From the Department of Biomedical Sciences, Creighton University School of Medicine, Omaha, Nebraska 68178.
Huizhan
H
Liu
From the Department of Biomedical Sciences, Creighton University School of Medicine, Omaha, Nebraska 68178.
Jie
J
Tang
From the Department of Biomedical Sciences, Creighton University School of Medicine, Omaha, Nebraska 68178.
Jason L
JL
Pecka
From the Department of Biomedical Sciences, Creighton University School of Medicine, Omaha, Nebraska 68178.
Marcus P D
MP
Hatfield
From the Department of Biomedical Sciences, Creighton University School of Medicine, Omaha, Nebraska 68178 kirkbeisel@creighton.edu.
Kirk W
KW
Beisel
D027322:Amino Acid Transport System X-AG / Q000378:metabolism*; D000818:Animals; D027321:Anion Transport Proteins / Q000737:chemistry*; D000838:Anions; D002942:Circular Dichroism; D018360:Crystallography, X-Ray; D004563:Electrochemistry; D004594:Electrophysiology; D005849:Gerbillinae; D057809:HEK293 Cells; D018072:Hair Cells, Auditory, Outer / Q000378:metabolism*; D006309:Hearing; D006801:Humans; D018613:Microscopy, Confocal; D056004:Molecular Dynamics Simulation; D016297:Mutagenesis, Site-Directed; D010455:Peptides / Q000737:chemistry; D011485:Protein Binding; D011487:Protein Conformation; D017510:Protein Folding; D043902:Pyrococcus horikoshii / Q000378:metabolism; D051381:Rats; D000076988:Sulfate Transporters; D014443:Tyrosine / Q000737:chemistry
D027322:Amino Acid Transport System X-AG; D027321:Anion Transport Proteins; D000838:Anions; D010455:Peptides; C498719:SLC26A5 protein, human; C409207:Slc26a5 protein, rat; D000076988:Sulfate Transporters; D014443:Tyrosine
anion-π interactions; electrophysiology; hair cell; homology modeling; molecular dynamics; prestin; protein structure; transmembrane domain