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.

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  • Additional Information
    • Source:
      Publisher: MDPI Country of Publication: Switzerland NLM ID: 101092791 Publication Model: Electronic Cited Medium: Internet ISSN: 1422-0067 (Electronic) Linking ISSN: 14220067 NLM ISO Abbreviation: Int J Mol Sci Subsets: MEDLINE
    • Publication Information:
      Original Publication: Basel, Switzerland : MDPI, [2000-
    • Subject Terms:
      Molecular Dynamics Simulation* ; Mutation* ; Protein Stability*; Amino Acid Transport Systems/*chemistry ; Antiporters/*chemistry ; Escherichia coli Proteins/*chemistry ; Large Neutral Amino Acid-Transporter 1/*metabolism; Amino Acid Transport Systems/genetics ; Amino Acid Transport Systems/metabolism ; Antiporters/genetics ; Antiporters/metabolism ; Escherichia coli Proteins/genetics ; Escherichia coli Proteins/metabolism ; Hot Temperature ; Humans ; Large Neutral Amino Acid-Transporter 1/chemistry ; Ligands ; Protein Binding ; Sequence Homology
    • Abstract:
      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 Escherichia coli, 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 ( T m 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 T m s indicated stabilization of AdiC variants upon ligand binding, in which T m 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.
      Competing Interests: The authors declare no conflict of interest.
    • References:
      Nucleic Acids Res. 2013 Jul;41(Web Server issue):W597-600. (PMID: 23671338)
      Biochem Pharmacol. 2017 Nov 1;143:39-52. (PMID: 28709952)
      Int J Biochem Cell Biol. 2015 Oct;67:25-33. (PMID: 26256001)
      Proc Natl Acad Sci U S A. 2013 Apr 2;110(14):5480-5. (PMID: 23509259)
      Mol Endocrinol. 2015 Jun;29(6):933-42. (PMID: 25945809)
      Proc Natl Acad Sci U S A. 2011 Mar 8;108(10):3935-40. (PMID: 21368142)
      Mol Cell Endocrinol. 2017 Dec 15;458:68-75. (PMID: 28341457)
      Bioorg Med Chem Lett. 2016 Jun 1;26(11):2616-2621. (PMID: 27106710)
      Curr Top Med Chem. 2013;13(7):843-56. (PMID: 23578028)
      Biochim Biophys Acta. 2017 Apr;1861(4):727-736. (PMID: 28088504)
      Nature. 2010 Feb 11;463(7282):828-32. (PMID: 20090677)
      Science. 2009 Jun 19;324(5934):1565-8. (PMID: 19478139)
      Nat Protoc. 2012 Sep;7(9):1569-78. (PMID: 22864198)
      Nat Protoc. 2016 Aug;11(8):1554-71. (PMID: 27466713)
      Nat Rev Cancer. 2016 Oct;16(10 ):619-34. (PMID: 27492215)
      Proc Natl Acad Sci U S A. 2014 Feb 25;111(8):2966-71. (PMID: 24516142)
      Nat Rev Microbiol. 2004 Nov;2(11):898-907. (PMID: 15494746)
      Biochem Pharmacol. 2013 Jun 1;85(11):1672-83. (PMID: 23567998)
      Proc Natl Acad Sci U S A. 2013 Apr 9;110(15):5893-7. (PMID: 23530220)
      Cell. 2009 Feb 6;136(3):521-34. (PMID: 19203585)
      J Biol Chem. 2007 Jan 5;282(1):176-82. (PMID: 17099215)
      Curr Protoc Bioinformatics. 2006 Oct;Chapter 5:Unit-5.6. (PMID: 18428767)
      Medchemcomm. 2016 Jun 1;7(6):1069-1081. (PMID: 27672436)
      J Biol Chem. 2005 May 13;280(19):19364-72. (PMID: 15757906)
      Proteins. 2009 Dec;77(4):778-95. (PMID: 19603484)
      Nucleic Acids Res. 2008 Apr;36(7):2295-300. (PMID: 18287115)
      Bioorg Med Chem Lett. 2016 Oct 15;26(20):5000-5006. (PMID: 27624080)
      Biophys J. 2014 Apr 15;106(8):1660-70. (PMID: 24739165)
      Proc Natl Acad Sci U S A. 2016 Sep 13;113(37):10358-63. (PMID: 27582465)
      J Biol Chem. 2008 Nov 28;283(48):33240-8. (PMID: 18819925)
      Biochemistry. 2012 Feb 28;51(8):1577-85. (PMID: 22304019)
      Mol Cell Endocrinol. 2017 Mar 5;443:163-174. (PMID: 28108384)
      Biol Chem. 2011 Jan;392(1-2):143-50. (PMID: 21194358)
      Nature. 2009 Aug 20;460(7258):1040-3. (PMID: 19578361)
      Mol Aspects Med. 2013 Apr-Jun;34(2-3):139-58. (PMID: 23506863)
      Methods. 2011 Dec;55(4):324-9. (PMID: 21840396)
    • Grant Information:
      R01 GM108911 United States GM NIGMS NIH HHS
    • Contributed Indexing:
      Keywords: ; ">l-arginine/agmatine transporter; ">l-type amino acid transporter; AdiC; LAT1; acid resistance; cancer metabolism; enterobacteria; melting temperature; thermostability
    • Accession Number:
      0 (AdiC protein, E coli)
      0 (Amino Acid Transport Systems)
      0 (Antiporters)
      0 (Escherichia coli Proteins)
      0 (Large Neutral Amino Acid-Transporter 1)
      0 (Ligands)
    • Publication Date:
      Date Created: 20180321 Date Completed: 20180905 Latest Revision: 20181114
    • Publication Date:
      20240829
    • Accession Number:
      PMC5877779
    • Accession Number:
      10.3390/ijms19030918
    • Accession Number:
      29558430