Interaction between the motor protein prestin and the transporter protein VAPA

Soma Sengupta, Katharine K. Miller, Kazuaki Homma, Roxanne Edge, Mary Ann Cheatham, Peter Dallos, Jing Zheng

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Prestin is the motor protein responsible for cochlear outer hair cell (OHC) somatic electromotility. Eliminating this abundant basolateral membrane protein not only causes loss of frequency selectivity and hearing sensitivity, but also leads to OHC death. A membrane-based yeast two-hybrid approach was used to screen an OHC-enriched cDNA (complementary Deoxyribonucleic Acid) library in order to identify prestin-associated proteins. Several proteins were recognized as potential prestin partners, including vesicle-associated membrane protein associated protein A (VAPA or VAP-33). VAPA is an integral membrane protein that plays an important role in membrane trafficking, endoplasmic reticulum homeostasis, and the stress-signaling system. The connection between VAPA and prestin was confirmed through co-immunoprecipitation experiments. This new finding prompted the investigation of the interaction between VAPA and prestin in outer hair cells. By comparing VAPA expression between wild-type OHCs and OHCs derived from prestin-knockout mice, we found that VAPA is expressed in OHCs and the quantity of VAPA expressed is related to the presence of prestin. In other words, less VAPA protein is found in OHCs lacking prestin. Thus, prestin appears to modify the expression of VAPA protein in OHCs. Intriguingly, more prestin protein appears at the plasma membrane when VAPA is co-expressed with prestin. These data suggest that VAPA could be involved in prestin's transportation inside OHCs and may facilitate the targeting of this abundant OHC protein to the plasma membrane.

Original languageEnglish (US)
Pages (from-to)796-804
Number of pages9
JournalBiochimica et Biophysica Acta - Molecular Cell Research
Volume1803
Issue number7
DOIs
StatePublished - Jul 1 2010

Fingerprint

Outer Auditory Hair Cells
Proteins
Membrane Proteins
R-SNARE Proteins
Cell Membrane
Endoplasmic Reticulum Stress
Membranes
Staphylococcal Protein A
Immunoprecipitation
Knockout Mice
Hearing
Homeostasis
Cell Death
Yeasts
DNA

Keywords

  • Outer hair cell
  • Prestin
  • Protein trafficking
  • VAPA

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

Sengupta, Soma ; Miller, Katharine K. ; Homma, Kazuaki ; Edge, Roxanne ; Cheatham, Mary Ann ; Dallos, Peter ; Zheng, Jing. / Interaction between the motor protein prestin and the transporter protein VAPA. In: Biochimica et Biophysica Acta - Molecular Cell Research. 2010 ; Vol. 1803, No. 7. pp. 796-804.
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Interaction between the motor protein prestin and the transporter protein VAPA. / Sengupta, Soma; Miller, Katharine K.; Homma, Kazuaki; Edge, Roxanne; Cheatham, Mary Ann; Dallos, Peter; Zheng, Jing.

In: Biochimica et Biophysica Acta - Molecular Cell Research, Vol. 1803, No. 7, 01.07.2010, p. 796-804.

Research output: Contribution to journalArticle

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T1 - Interaction between the motor protein prestin and the transporter protein VAPA

AU - Sengupta, Soma

AU - Miller, Katharine K.

AU - Homma, Kazuaki

AU - Edge, Roxanne

AU - Cheatham, Mary Ann

AU - Dallos, Peter

AU - Zheng, Jing

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AB - Prestin is the motor protein responsible for cochlear outer hair cell (OHC) somatic electromotility. Eliminating this abundant basolateral membrane protein not only causes loss of frequency selectivity and hearing sensitivity, but also leads to OHC death. A membrane-based yeast two-hybrid approach was used to screen an OHC-enriched cDNA (complementary Deoxyribonucleic Acid) library in order to identify prestin-associated proteins. Several proteins were recognized as potential prestin partners, including vesicle-associated membrane protein associated protein A (VAPA or VAP-33). VAPA is an integral membrane protein that plays an important role in membrane trafficking, endoplasmic reticulum homeostasis, and the stress-signaling system. The connection between VAPA and prestin was confirmed through co-immunoprecipitation experiments. This new finding prompted the investigation of the interaction between VAPA and prestin in outer hair cells. By comparing VAPA expression between wild-type OHCs and OHCs derived from prestin-knockout mice, we found that VAPA is expressed in OHCs and the quantity of VAPA expressed is related to the presence of prestin. In other words, less VAPA protein is found in OHCs lacking prestin. Thus, prestin appears to modify the expression of VAPA protein in OHCs. Intriguingly, more prestin protein appears at the plasma membrane when VAPA is co-expressed with prestin. These data suggest that VAPA could be involved in prestin's transportation inside OHCs and may facilitate the targeting of this abundant OHC protein to the plasma membrane.

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