Molecular origin of plasma membrane citrate transporter in human prostate epithelial cells

MacIej P. Mazurek, Puttur D. Prasad, Elangovan Gopal, Scott P. Fraser, Leszek Bolt, Nahit Rizaner, Christopher P. Palmer, Christopher S. Foster, Ferdinando Palmieri, Vadivel Ganapathy, Walter Stühmer, Mustafa B.A. Djamgoz, Maria E. Mycielska

Research output: Contribution to journalArticle

16 Scopus citations

Abstract

The prostate is a highly specialized mammalian organ that produces and releases large amounts of citrate. However, the citrate release mechanism is not known. Here, we present the results of molecular cloning of a citrate transporter from human normal prostate epithelial PNT2-C2 cells shown previously to express such a mechanism. By using rapid amplification of cDNA ends PCR, we determined that the prostatic carrier is an isoform of the mitochondrial transporter SLC25A1 with a different first exon. We confirmed the functionality of the clone by expressing it in human embryonic kidney cells and performing radiotracer experiments and whole-cell patch-clamp recordings. By using short interfering RNAs targeting different parts of the sequence, we confirmed that the cloned protein is the main prostatic transporter responsible for citrate release. We also produced a specific antibody and localized the cloned transporter protein to the plasma membrane of the cells. By using the same antibody, we have shown that the cloned transporter is expressed in non-malignant human tissues.

Original languageEnglish (US)
Pages (from-to)431-437
Number of pages7
JournalEMBO Reports
Volume11
Issue number6
DOIs
StatePublished - Jun 1 2010

Keywords

  • Citrate
  • Prostate
  • Release
  • SLC25
  • Transporter

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Genetics

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  • Cite this

    Mazurek, M. P., Prasad, P. D., Gopal, E., Fraser, S. P., Bolt, L., Rizaner, N., Palmer, C. P., Foster, C. S., Palmieri, F., Ganapathy, V., Stühmer, W., Djamgoz, M. B. A., & Mycielska, M. E. (2010). Molecular origin of plasma membrane citrate transporter in human prostate epithelial cells. EMBO Reports, 11(6), 431-437. https://doi.org/10.1038/embor.2010.51