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Molecular origin of plasma membrane citrate transporter in human prostate epithelial cells
Author(s) -
Mazurek Maciej P,
Prasad Puttur D,
Gopal Elangovan,
Fraser Scott P,
Bolt Leszek,
Rizaner Nahit,
Palmer Christopher P,
Foster Christopher S,
Palmieri Ferdinando,
Ganapathy Vadivel,
Stühmer Walter,
Djamgoz Mustafa B A,
Mycielska Maria E
Publication year - 2010
Publication title -
embo reports
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 4.584
H-Index - 184
eISSN - 1469-3178
pISSN - 1469-221X
DOI - 10.1038/embor.2010.51
Subject(s) - transporter , biology , microbiology and biotechnology , complementary dna , cloning (programming) , solute carrier family , hek 293 cells , kidney , prostate , clone (java method) , transport protein , biochemistry , gene , endocrinology , genetics , cancer , computer science , programming language
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.