4pd5 › Citations

Structural basis of nucleoside and nucleoside drug selectivity by concentrative nucleoside transporters.

<div class='caption-title'>(A) The vcCNT-7C8C trimer viewed from within the plane of the membrane. The location of the membrane is marked by rectangles. The scaffold domain of one protomer is colored light blue, and the transport domain is colored red, blue, orange, cyan, wheat, and brown. The other two protomers are colored gray. Uridine is shown bound to each protomer in stick representation. The nucleoside-binding site is delineated with dashed lines. vcCNT-7C8C functions similarly to wild type (Figure 1—figure supplement 1). (B) The vcCNT-7C8C protomer. The structure is rotated 120° about the trimer axis relative to A, zoomed in, and the other two protomers have been removed for clarity. (C) Nucleoside-binding site. Amino acid residues that interact with the uridine are labeled and shown in stick representation and were used for sequence identity calculation with hCNTs. Hydrogen bonds are shown as dashed lines. The uracil base is marked with a blue box, and the ribose is marked with a gray box. For a stereo view of the electron density in the nucleoside-binding site, see Figure 1—figure supplement 2. (D) Fluorescence titration of vcCNT with uridine. Uridine was titrated into solution containing vcCNT and the fluorescent nucleoside pyrrolo-cytidine, anisotropy was measured, and data were fit to a single-site competitive binding model to obtain a KD of 36 ± 3 μM (mean ± SEM, n = 3 measurements).</div>
<div class='caption-title'>Wild-type (WT) and 7C8C vcCNT were reconstituted into lipid vesicles and assessed for their ability to support the uptake of 2 μM of 3H-labeled nucleosides as described (Johnson et al., 2012). The data shown represent 2 min timepoints. Empty vesicles were included as a negative control.</div>
<div class='caption-title'>The nucleoside-binding site of vcCNT-7C8C bound to uridine is shown in stereo in stick representation in the same orientation and colored as in Figure 1C. Uridine is yellow and the red spheres are water molecules. The resolution is 2.1 Å and density shown is from a 2Fo–Fc electron density map contoured at 1σ.</div>
Johnson ZL, Lee JH, Lee K, Lee M, Kwon DY, Hong J, Lee SY
OpenAccess logo Elife 3 e03604 (2014)
Related entries: 4pb1, 4pb2, 4pd6, 4pd7, 4pd8, 4pd9, 4pda

Cited: 31 times
EuropePMC logo PMID: 25082345

Abstract

Concentrative nucleoside transporters (CNTs) are responsible for cellular entry of nucleosides, which serve as precursors to nucleic acids and act as signaling molecules. CNTs also play a crucial role in the uptake of nucleoside-derived drugs, including anticancer and antiviral agents. Understanding how CNTs recognize and import their substrates could not only lead to a better understanding of nucleoside-related biological processes but also the design of nucleoside-derived drugs that can better reach their targets. Here, we present a combination of X-ray crystallographic and equilibrium-binding studies probing the molecular origins of nucleoside and nucleoside drug selectivity of a CNT from Vibrio cholerae. We then used this information in chemically modifying an anticancer drug so that it is better transported by and selective for a single human CNT subtype. This work provides proof of principle for utilizing transporter structural and functional information for the design of compounds that enter cells more efficiently and selectively.

Reviews - 4pd5 mentioned but not cited (2)

  1. Elevator-type mechanisms of membrane transport. Garaeva AA, Slotboom DJ. Biochem Soc Trans 48 1227-1241 (2020)
  2. Toward a Molecular Basis of Cellular Nucleoside Transport in Humans. Wright NJ, Lee SY. Chem Rev 121 5336-5358 (2021)

Articles - 4pd5 mentioned but not cited (2)



Reviews citing this publication (7)

  1. SLC transporters as therapeutic targets: emerging opportunities. Lin L, Yee SW, Kim RB, Giacomini KM. Nat Rev Drug Discov 14 543-560 (2015)
  2. Multifaceted roles of extracellular DNA in bacterial physiology. Vorkapic D, Pressler K, Schild S. Curr Genet 62 71-79 (2016)
  3. Polypharmacology of conformationally locked methanocarba nucleosides. Jacobson KA, Tosh DK, Toti KS, Ciancetta A. Drug Discov Today 22 1782-1791 (2017)
  4. Recent advances on the inhibition of human solute carriers: Therapeutic implications and mechanistic insights. Wright NJ, Lee SY. Curr Opin Struct Biol 74 102378 (2022)
  5. An Analysis of Mechanisms for Cellular Uptake of miRNAs to Enhance Drug Delivery and Efficacy in Cancer Chemoresistance. Grixti JM, Ayers D, Day PJR. Noncoding RNA 7 27 (2021)
  6. Disruption of small molecule transporter systems by Transporter-Interfering Chemicals (TICs). Nicklisch SCT, Hamdoun A. FEBS Lett 594 4158-4185 (2020)
  7. Novel molecular mechanisms of doxorubicin cardiotoxicity: latest leading-edge advances and clinical implications. Robert Li Y, Traore K, Zhu H. Mol Cell Biochem (2023)

Articles citing this publication (20)



Quick links