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Interaction of TM4SF4 with the human thiamine (vitamin B1) transporter‐2 ( hTHTR‐2 ) in human intestinal epithelial cells (896.1)
Author(s) -
Subramanian Veedamali,
Nabokina Svetlana,
Said Hamid
Publication year - 2014
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.28.1_supplement.896.1
Subject(s) - thiamine , transporter , microbiology and biotechnology , biology , transmembrane protein , transport protein , function (biology) , biochemistry , chemistry , gene , receptor
The human thiamine transporter‐2 (hTHTR‐2) is involved in intestinal absorption of thiamine. Recent studies with membrane transporters of other nutrients have shown that these transporters have associated proteins that affect different aspects of their physiology and cell biology. Nothing is currently known about protein(s) that may interact with hTHTR‐2 in intestinal cells and influence its physiological function and cell biology.We addressed this issue using the yeast split‐ubiquitin two‐hybrid approach to screen a human intestinal cDNA library with the full‐length hTHTR‐2 as bait. We also performed GST‐pull‐down assay and co‐localization to confirm the existence of protein‐protein interaction. Our screening results have identified the human tetraspanin (transmembrane 4 super family member 4, TM4SF4) protein (which is abundantly expressed in the intestine) as a potential interactor with hTHTR‐2. The interaction between hTHTR‐2 and TM4SF4 were further confirmed by in vitro GST‐pull‐down assay and by live‐cell confocal imaging of HuTu‐80 cells co‐expressing hTHTR‐2‐GFP and Cherry‐TM4SF4. Co‐expression of hTHTR‐2 with TM4SF4 in HuTu‐80 cells led to a significant induction in hTHTR‐2 ‐ mediated thiamine uptake. These results show for the first time the existence of an accessory protein that interacts with hTHTR‐2 and influence its physiological function. (Supported by NIH grant DK 56061‐15). Grant Funding Source : NIH‐NIDDK