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2015 ; 10
(9
): e0136779
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The Role of Flexible Loops in Folding, Trafficking and Activity of Equilibrative
Nucleoside Transporters
#MMPMID26406980
Aseervatham J
; Tran L
; Machaca K
; Boudker O
PLoS One
2015[]; 10
(9
): e0136779
PMID26406980
show ga
Equilibrative nucleoside transporters (ENTs) are integral membrane proteins,
which reside in plasma membranes of all eukaryotic cells and mediate
thermodynamically downhill transport of nucleosides. This process is essential
for nucleoside recycling, and also plays a key role in terminating
adenosine-mediated cellular signaling. Furthermore, ENTs mediate the uptake of
many drugs, including anticancer and antiviral nucleoside analogues. The
structure and mechanism, by which ENTs catalyze trans-membrane transport of their
substrates, remain unknown. To identify the core of the transporter needed for
stability, activity, and for its correct trafficking to the plasma membrane, we
have expressed human ENT deletion mutants in Xenopus laevis oocytes and
determined their localization, transport properties and susceptibility to
inhibition. We found that the carboxyl terminal trans-membrane segments are
essential for correct protein folding and trafficking. In contrast, the soluble
extracellular and intracellular loops appear to be dispensable, and must be
involved in the fine-tuning of transport regulation.