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Broad range neutral amino acid transporter (B 0 AT1) requires association with TMEM27 for surface expression in renal cells
Author(s) -
Torrente Marta,
Arps Lisa,
Guetg Adriano,
Camargo Simone M.R.,
Verrey François
Publication year - 2012
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.26.1_supplement.1068.19
Subject(s) - angiotensin ii receptor type 1 , amino acid , chemistry , angiotensin ii , renin–angiotensin system , microbiology and biotechnology , in vivo , glycine , biochemistry , biology , receptor , endocrinology , genetics , blood pressure
TMEM27 (collectrin), a homologue of angiotensin converting enzyme 2 (ACE2), was previously shown in renal proximal tubule to regulate amino acid uptake via interaction with the broad range neutral amino acid transporter B 0 AT1, whereas ACE2 was shown to associate with B 0 AT1 in small intestine. To investigate the role of collectrin interaction with B 0 AT1, we generated Madin‐Darby canine kidney (MDCK) cell lines constitutively expressing B 0 AT1 and observed that this expression was not tolerated. Therefore, using a KRAB repressor system and lentiviral transduction, we produced cells that inducibly express either B 0 AT1, collectrin or both. As in vivo , collectrin was shown to be required for efficient B 0 AT1 cell surface expression and function. Furthermore, we demonstrated that collectrin co‐expression increases B 0 AT1 protein stability. Interestingly, expression levels of both B 0 AT1 and collectrin were increased when amino acid concentrations in the culture medium were reduced to physiological levels. In conclusion, we generated cell lines expressing inducible B 0 AT1 and collectrin recapitulating the dependence of B 0 AT1 expression on collectrin co‐expression and thus representing a very good model for testing the role and the mechanism of B 0 AT1‐collectrin interaction and the impact of the amino acid concentration for the control of its expression.