Identification of a novel sodium-coupled oligopeptide transporter (SOPT2) in mouse and human retinal pigment epithelial cells

Paresh P. Chothe, Santoshanand V. Thakkar, Jaya P. Gnana-Prakasam, Sudha Ananth, David R. Hinton, Ram Kannan, Sylvia B. Smith, Pamela M. Martin, Vadivel Ganapathy

Research output: Contribution to journalArticle

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Abstract

PURPOSE. A sodium-coupled oligopeptide transporter (SOPT1) was described originally in ARPE-19 cells. The transporter is inducible by HIV-1 Tat. Recent studies of conjunctival epithelial cells have identified a second oligopeptide transporter (SOPT2). This study was conducted to determine whether the newly discovered SOPT2 is expressed in ARPE-19 cells, to examine whether the new transporter is also inducible by HIV-1 Tat, and to find out whether this transporter is expressed in primary RPE cells. METHODS. The transport activity of SOPT2 was monitored in control and Tat-expressing ARPE-19 cells and in primary mouse and human fetal RPE cells by the uptake of the synthetic opioid peptide DADLE ((H-Tyr-D-Ala-Gly-Phe-D-Leu-OH) and by its susceptibility to inhibition by small peptides. Substrate selectivity was examined by competition studies and kinetic parameters were determined by saturation analysis. RESULTS. ARPE-19 cells express DADLE uptake activity that is inhibited by small peptides, indicating expression of SOPT2 in these cells. The activity of SOPT2 is induced by HIV-1 Tat. SOPT2 accepts endogenous and synthetic opioid peptides as substrates, but nonpeptide opiate antagonists are excluded. An 11-amino-acid HIV-1 Tat peptide also serves as a high-affinity substrate for the transporter. Primary cultures of mouse and human fetal RPE cells express SOPT2. The transporter is partially Na +-dependent with comparable substrate selectivity and inhibitor specificity in the presence and absence of Na +. CONCLUSIONS. ARPE-19 cells as well as primary mouse and human fetal RPE cells express the newly discovered oligopeptide transporter SOPT2, and the transporter is induced by HIV-1 Tat in ARPE-19 cells.

Original languageEnglish (US)
Pages (from-to)413-420
Number of pages8
JournalInvestigative Ophthalmology and Visual Science
Volume51
Issue number1
DOIs
StatePublished - Jan 1 2010

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Oligopeptides
Retinal Pigments
Epithelial Cells
Sodium
HIV-1
Leucine-2-Alanine Enkephalin
Opioid Peptides
Peptides
Opiate Alkaloids
Artificial Cells
Amino Acids

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

Cite this

Chothe, P. P., Thakkar, S. V., Gnana-Prakasam, J. P., Ananth, S., Hinton, D. R., Kannan, R., ... Ganapathy, V. (2010). Identification of a novel sodium-coupled oligopeptide transporter (SOPT2) in mouse and human retinal pigment epithelial cells. Investigative Ophthalmology and Visual Science, 51(1), 413-420. https://doi.org/10.1167/iovs.09-4048

Identification of a novel sodium-coupled oligopeptide transporter (SOPT2) in mouse and human retinal pigment epithelial cells. / Chothe, Paresh P.; Thakkar, Santoshanand V.; Gnana-Prakasam, Jaya P.; Ananth, Sudha; Hinton, David R.; Kannan, Ram; Smith, Sylvia B.; Martin, Pamela M.; Ganapathy, Vadivel.

In: Investigative Ophthalmology and Visual Science, Vol. 51, No. 1, 01.01.2010, p. 413-420.

Research output: Contribution to journalArticle

Chothe, Paresh P. ; Thakkar, Santoshanand V. ; Gnana-Prakasam, Jaya P. ; Ananth, Sudha ; Hinton, David R. ; Kannan, Ram ; Smith, Sylvia B. ; Martin, Pamela M. ; Ganapathy, Vadivel. / Identification of a novel sodium-coupled oligopeptide transporter (SOPT2) in mouse and human retinal pigment epithelial cells. In: Investigative Ophthalmology and Visual Science. 2010 ; Vol. 51, No. 1. pp. 413-420.
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T1 - Identification of a novel sodium-coupled oligopeptide transporter (SOPT2) in mouse and human retinal pigment epithelial cells

AU - Chothe, Paresh P.

AU - Thakkar, Santoshanand V.

AU - Gnana-Prakasam, Jaya P.

AU - Ananth, Sudha

AU - Hinton, David R.

AU - Kannan, Ram

AU - Smith, Sylvia B.

AU - Martin, Pamela M.

AU - Ganapathy, Vadivel

PY - 2010/1/1

Y1 - 2010/1/1

N2 - PURPOSE. A sodium-coupled oligopeptide transporter (SOPT1) was described originally in ARPE-19 cells. The transporter is inducible by HIV-1 Tat. Recent studies of conjunctival epithelial cells have identified a second oligopeptide transporter (SOPT2). This study was conducted to determine whether the newly discovered SOPT2 is expressed in ARPE-19 cells, to examine whether the new transporter is also inducible by HIV-1 Tat, and to find out whether this transporter is expressed in primary RPE cells. METHODS. The transport activity of SOPT2 was monitored in control and Tat-expressing ARPE-19 cells and in primary mouse and human fetal RPE cells by the uptake of the synthetic opioid peptide DADLE ((H-Tyr-D-Ala-Gly-Phe-D-Leu-OH) and by its susceptibility to inhibition by small peptides. Substrate selectivity was examined by competition studies and kinetic parameters were determined by saturation analysis. RESULTS. ARPE-19 cells express DADLE uptake activity that is inhibited by small peptides, indicating expression of SOPT2 in these cells. The activity of SOPT2 is induced by HIV-1 Tat. SOPT2 accepts endogenous and synthetic opioid peptides as substrates, but nonpeptide opiate antagonists are excluded. An 11-amino-acid HIV-1 Tat peptide also serves as a high-affinity substrate for the transporter. Primary cultures of mouse and human fetal RPE cells express SOPT2. The transporter is partially Na +-dependent with comparable substrate selectivity and inhibitor specificity in the presence and absence of Na +. CONCLUSIONS. ARPE-19 cells as well as primary mouse and human fetal RPE cells express the newly discovered oligopeptide transporter SOPT2, and the transporter is induced by HIV-1 Tat in ARPE-19 cells.

AB - PURPOSE. A sodium-coupled oligopeptide transporter (SOPT1) was described originally in ARPE-19 cells. The transporter is inducible by HIV-1 Tat. Recent studies of conjunctival epithelial cells have identified a second oligopeptide transporter (SOPT2). This study was conducted to determine whether the newly discovered SOPT2 is expressed in ARPE-19 cells, to examine whether the new transporter is also inducible by HIV-1 Tat, and to find out whether this transporter is expressed in primary RPE cells. METHODS. The transport activity of SOPT2 was monitored in control and Tat-expressing ARPE-19 cells and in primary mouse and human fetal RPE cells by the uptake of the synthetic opioid peptide DADLE ((H-Tyr-D-Ala-Gly-Phe-D-Leu-OH) and by its susceptibility to inhibition by small peptides. Substrate selectivity was examined by competition studies and kinetic parameters were determined by saturation analysis. RESULTS. ARPE-19 cells express DADLE uptake activity that is inhibited by small peptides, indicating expression of SOPT2 in these cells. The activity of SOPT2 is induced by HIV-1 Tat. SOPT2 accepts endogenous and synthetic opioid peptides as substrates, but nonpeptide opiate antagonists are excluded. An 11-amino-acid HIV-1 Tat peptide also serves as a high-affinity substrate for the transporter. Primary cultures of mouse and human fetal RPE cells express SOPT2. The transporter is partially Na +-dependent with comparable substrate selectivity and inhibitor specificity in the presence and absence of Na +. CONCLUSIONS. ARPE-19 cells as well as primary mouse and human fetal RPE cells express the newly discovered oligopeptide transporter SOPT2, and the transporter is induced by HIV-1 Tat in ARPE-19 cells.

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