The C-terminus of prestin influences nonlinear capacitance and plasma membrane targeting

Jing Zheng*, Guo Guang Du, Keiji Matsuda, Alex Orem, Sal Aguiñaga, Levente Deák, Enrique Navarrete, Laird D. Madison, Peter Dallos

*Corresponding author for this work

Research output: Contribution to journalArticle

48 Citations (Scopus)

Abstract

Prestin is a unique molecular-motor protein expressed in the lateral plasma membrane of outer hair cells (OHC) in the organ of Corti of the mammalian cochlea. It is thought that prestin undergoes conformational changes driven by the cell's membrane potential. The resulting alterations in OHC-length are assumed to constitute the cochlear amplifier. Prestin is a member of the anion solute carrier family 26 (SCL26A), but it is different from other family members in its unique function of voltage-driven motility. Because the C-terminus is the least conserved region in the family, we investigated its influence with a series of deletion, point and chimeric mutants. The function and cellular expression of mutants were examined in a heterologous expression system by measurement of nonlinear capacitance (NLC) and immunofluorescence. Each mutant produced a unique mixture of patterns of cell morphologies, which were classified as to the location of prestin within the cell. The data from deletion mutants (Del516, Del525, Del630, Del590, Del709, Del719) revealed that nearly the full length (>708 amino acids) of the protein was required for normal prestin expression and function. Since most deletion mutations eliminated plasma membrane targeting, chimeric proteins were constructed by fusing prestin, at amino acid 515 or 644, with the homologous portion of the C-terminus from the two most closely related SLC26A members, pendrin and putative anion exchanger 1. These chimeric proteins were again improperly (but differently) targeted than simple truncation mutants, and all lacked functional phenotype. When two of the potential basolateral membrane-targeting motifs were mutated (Y520A/Y526A), incomplete plasma membrane expression was seen. We also show that some double point mutations (V499G/Y501H) fully express in the plasma membrane but lack NLC. These non-charged amino acids may have unrevealed important roles in prestin's function. Together, these data suggest that certain specific sequences and individual amino acids in the C-terminus are necessary for correct cellular distribution and function.

Original languageEnglish (US)
Pages (from-to)2987-2996
Number of pages10
JournalJournal of cell science
Volume118
Issue number13
DOIs
StatePublished - Jul 1 2005

Fingerprint

Outer Auditory Hair Cells
Cell Membrane
Cochlea
Amino Acids
Membrane Potentials
Molecular Motor Proteins
Erythrocyte Anion Exchange Protein 1
Organ of Corti
Sequence Deletion
Protein Transport
Point Mutation
Fluorescent Antibody Technique
Anions
Amino Acid Sequence
Proteins
Phenotype

Keywords

  • C-terminus
  • Membrane targeting
  • Nonlinear capacitance
  • Outer hair cells
  • Prestin
  • STAS

ASJC Scopus subject areas

  • Cell Biology

Cite this

Zheng, J., Du, G. G., Matsuda, K., Orem, A., Aguiñaga, S., Deák, L., ... Dallos, P. (2005). The C-terminus of prestin influences nonlinear capacitance and plasma membrane targeting. Journal of cell science, 118(13), 2987-2996. https://doi.org/10.1242/jcs.02431
Zheng, Jing ; Du, Guo Guang ; Matsuda, Keiji ; Orem, Alex ; Aguiñaga, Sal ; Deák, Levente ; Navarrete, Enrique ; Madison, Laird D. ; Dallos, Peter. / The C-terminus of prestin influences nonlinear capacitance and plasma membrane targeting. In: Journal of cell science. 2005 ; Vol. 118, No. 13. pp. 2987-2996.
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Zheng, J, Du, GG, Matsuda, K, Orem, A, Aguiñaga, S, Deák, L, Navarrete, E, Madison, LD & Dallos, P 2005, 'The C-terminus of prestin influences nonlinear capacitance and plasma membrane targeting', Journal of cell science, vol. 118, no. 13, pp. 2987-2996. https://doi.org/10.1242/jcs.02431

The C-terminus of prestin influences nonlinear capacitance and plasma membrane targeting. / Zheng, Jing; Du, Guo Guang; Matsuda, Keiji; Orem, Alex; Aguiñaga, Sal; Deák, Levente; Navarrete, Enrique; Madison, Laird D.; Dallos, Peter.

In: Journal of cell science, Vol. 118, No. 13, 01.07.2005, p. 2987-2996.

Research output: Contribution to journalArticle

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T1 - The C-terminus of prestin influences nonlinear capacitance and plasma membrane targeting

AU - Zheng, Jing

AU - Du, Guo Guang

AU - Matsuda, Keiji

AU - Orem, Alex

AU - Aguiñaga, Sal

AU - Deák, Levente

AU - Navarrete, Enrique

AU - Madison, Laird D.

AU - Dallos, Peter

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KW - Membrane targeting

KW - Nonlinear capacitance

KW - Outer hair cells

KW - Prestin

KW - STAS

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