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Identification of the multivalent PDZ domain protein PDZK1 as a binding partner of sodium‐coupled monocarboxylate cotransporter 2 (SMCT2) by yeast two‐hybrid assay
Author(s) -
Srivastava Sunena,
Anzai Naohiko,
Yamanishi Ai,
Ganapathy Vadivel,
Endou Hitoshi
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.1_supplement.1204.3
Subject(s) - pdz domain , cotransporter , yeast , complementary dna , cdna library , two hybrid screening , cytoplasm , saccharomyces cerevisiae , symporter , transporter , biochemistry , monocarboxylate transporter , peptide sequence , transport protein , chemistry , biology , microbiology and biotechnology , sodium , gene , organic chemistry
Sodium‐coupled monocarboxylate transporter SMCT2 ( SLC5A12 ), mediates the transport of pyruvate, nicotinate and lactate (Gopal et al., BBA. 2007). Since URAT1 transports urate in exchange for intracellular organic anions such as lactate and nicotinate, SMCT2, as well as SMCT1, seems important for the transport function of URAT1 (Anzai et al. Curr Opin Rheumatol, 2007). In addition, SMCT2 C‐terminal domain is in the cytoplasm and contains PDZ motif, suggesting that it interacts with PDZ proteins. In the present study, we used the yeast two‐hybrid screening to investigate the putative SMCT2‐associated proteins in the kidney. Using the SMCT2 C‐terminus (SMCT2‐CT) as bait, we performed a yeast two‐hybrid screen of a human adult kidney cDNA library. From a total of 1.2 [~ 10 7 independent colonies screened, 34 positive clones were obtained. Of these, 8 yielded an identical sequence encoding the gene for the multivalent PDZ protein PDZK1. Deletion of C‐terminal PDZ motif abolished the interaction with PDZK1 in the yeast two‐hybrid system. In addition, the first, second and fourth PDZ domains of PDZK1 associate with SMCT2‐CT. The elucidation of these interactions may lead to the further understanding of functional regulation of urate transport via monocarboxylate handling in human kidney.