title: Molecular origin of plasma membrane citrate transporter in human prostate epithelial cells.
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.
authors.affiliation: Division of Cell and Molecular Biology, Sir Alexander Fleming Building, Imperial College London, South Kensington Campus, London SW7 2AZ, UK.
authors.forename: Maciej P
authors.initials: MP
authors.lastname: Mazurek
authors.forename: Puttur D
authors.initials: PD
authors.lastname: Prasad
authors.forename: Elangovan
authors.initials: E
authors.lastname: Gopal
authors.forename: Scott P
authors.initials: SP
authors.lastname: Fraser
authors.forename: Leszek
authors.initials: L
authors.lastname: Bolt
authors.forename: Nahit
authors.initials: N
authors.lastname: Rizaner
authors.forename: Christopher P
authors.initials: CP
authors.lastname: Palmer
authors.forename: Christopher S
authors.initials: CS
authors.lastname: Foster
authors.forename: Ferdinando
authors.initials: F
authors.lastname: Palmieri
authors.forename: Vadivel
authors.initials: V
authors.lastname: Ganapathy
authors.forename: Walter
authors.initials: W
authors.lastname: Stühmer
authors.forename: Mustafa B A
authors.initials: MB
authors.lastname: Djamgoz
authors.forename: Maria E
authors.initials: ME
authors.lastname: Mycielska
mesh_terms: D000595:Amino Acid Sequence; D027321:Anion Transport Proteins / Q000737:chemistry / Q000378:metabolism; D001692:Biological Transport; D002352:Carrier Proteins / Q000737:chemistry / Q000378:metabolism*; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism*; D019343:Citric Acid / Q000378:metabolism; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism*; D019143:Evolution, Molecular; D020868:Gene Silencing; D006801:Humans; D007150:Immunohistochemistry; D007477:Ions / Q000378:metabolism; D008297:Male; D024101:Mitochondrial Proteins / Q000737:chemistry / Q000378:metabolism; D008969:Molecular Sequence Data; D027361:Organic Anion Transporters; D011467:Prostate / Q000166:cytology*; D020033:Protein Isoforms / Q000737:chemistry / Q000378:metabolism
chemical_list: D027321:Anion Transport Proteins; D002352:Carrier Proteins; D007477:Ions; D024101:Mitochondrial Proteins; D027361:Organic Anion Transporters; D020033:Protein Isoforms; C532592:Slc25a1 protein, human; C020723:citrate-binding transport protein; D019343:Citric Acid
references.citation: DNA Seq. 1997;7(3-4):127-39
references.pmid: 9254007
references.citation: J Biol Chem. 2002 Oct 18;277(42):39469-76
references.pmid: 12177002
references.citation: Horm Metab Res. 2002 Aug;34(8):417-24
references.pmid: 12198595
references.citation: Eur J Radiol. 2007 Sep;63(3):351-60
references.pmid: 17709223
references.citation: Biochim Biophys Acta. 1989 Nov 23;977(2):171-6
references.pmid: 2804096
references.citation: Biochim Biophys Acta. 1993 Apr 5;1142(1-2):139-45
references.pmid: 8457581
references.citation: Clin Cancer Res. 2005 Aug 1;11(15):5381-9
references.pmid: 16061851
references.citation: Prostate. 1994;24(3):139-42
references.pmid: 8115279
references.citation: Biochem J. 2009 Apr 15;419(2):369-75
references.pmid: 19143589
references.citation: Mol Reprod Dev. 2006 Mar;73(3):369-78
references.pmid: 16362974
references.citation: Plant Physiol. 1999 Dec;121(4):1375-82
references.pmid: 10594125
references.citation: J Physiol. 2005 Mar 1;563(Pt 2):393-408
references.pmid: 15611019
references.citation: Breast Cancer Res Treat. 2007 Jan;101(2):149-60
references.pmid: 16838113
references.citation: J Physiol. 2004 Sep 15;559(Pt 3):821-33
references.pmid: 15254154
references.citation: FEBS Lett. 1995 Jan 9;357(3):297-300
references.pmid: 7835431