Effects of Mutations and Ligands on the Thermostability of the l-Arginine/Agmatine Antiporter AdiC and Deduced Insights into Ligand-Binding of Human l-Type Amino Acid Transporters.

Ilgü, Hüseyin; Jeckelmann, Jean-Marc; Colas, Claire; Ucurum Fotiadis, Zöhre; Schlessinger, Avner; Fotiadis, Dimitrios José (2018). Effects of Mutations and Ligands on the Thermostability of the l-Arginine/Agmatine Antiporter AdiC and Deduced Insights into Ligand-Binding of Human l-Type Amino Acid Transporters. International journal of molecular sciences, 19(3) Molecular Diversity Preservation International MDPI 10.3390/ijms19030918

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The l-arginine/agmatine transporter AdiC is a prokaryotic member of the SLC7 family, which enables pathogenic enterobacteria to survive the extremely acidic gastric environment. Wild-type AdiC from as well as its previously reported point mutants N22A and S26A, were overexpressed homologously and purified to homogeneity. A size-exclusion chromatography-based thermostability assay was used to determine the melting temperatures (s) of the purified AdiC variants in the absence and presence of the selected ligands l-arginine (Arg), agmatine, l-arginine methyl ester, and l-arginine amide. The resulting s indicated stabilization of AdiC variants upon ligand binding, in which s and ligand binding affinities correlated positively. Considering results from this and previous studies, we revisited the role of AdiC residue S26 in Arg binding and proposed interactions of the α-carboxylate group of Arg exclusively with amide groups of the AdiC backbone. In the context of substrate binding in the human SLC7 family member l-type amino acid transporter-1 (LAT1; SLC7A5), an analogous role of S66 in LAT1 to S26 in AdiC is discussed based on homology modeling and amino acid sequence analysis. Finally, we propose a binding mechanism for l-amino acid substrates to LATs from the SLC7 family.

Item Type:

Journal Article (Original Article)

Division/Institute:

04 Faculty of Medicine > Faculty Institutions > NCCR TransCure
04 Faculty of Medicine > Pre-clinic Human Medicine > Institute of Biochemistry and Molecular Medicine

UniBE Contributor:

Ilgü, Hüseyin; Jeckelmann, Jean-Marc; Ucurum Fotiadis, Zöhre and Fotiadis, Dimitrios José

Subjects:

500 Science > 570 Life sciences; biology
600 Technology > 610 Medicine & health

ISSN:

1661-6596

Publisher:

Molecular Diversity Preservation International MDPI

Language:

English

Submitter:

Barbara Järmann-Bangerter

Date Deposited:

27 Feb 2019 10:37

Last Modified:

01 Nov 2019 06:41

Publisher DOI:

10.3390/ijms19030918

PubMed ID:

29558430

Uncontrolled Keywords:

">l-arginine/agmatine transporter ">l-type amino acid transporter AdiC LAT1 acid resistance cancer metabolism enterobacteria melting temperature thermostability

BORIS DOI:

10.7892/boris.124491

URI:

https://boris.unibe.ch/id/eprint/124491

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