Structural basis for mammalian nucleotide sugar transport

Elife. 2019 Apr 15;8:e45221. doi: 10.7554/eLife.45221.

Abstract

Nucleotide-sugar transporters (NSTs) are critical components of the cellular glycosylation machinery. They transport nucleotide-sugar conjugates into the Golgi lumen, where they are used for the glycosylation of proteins and lipids, and they then subsequently transport the nucleotide monophosphate byproduct back to the cytoplasm. Dysregulation of human NSTs causes several debilitating diseases, and NSTs are virulence factors for many pathogens. Here we present the first crystal structures of a mammalian NST, the mouse CMP-sialic acid transporter (mCST), in complex with its physiological substrates CMP and CMP-sialic acid. Detailed visualization of extensive protein-substrate interactions explains the mechanisms governing substrate selectivity. Further structural analysis of mCST's unique lumen-facing partially-occluded conformation, coupled with the characterization of substrate-induced quenching of mCST's intrinsic tryptophan fluorescence, reveals the concerted conformational transitions that occur during substrate transport. These results provide a framework for understanding the effects of disease-causing mutations and the mechanisms of this diverse family of transporters.

Keywords: Golgi; glycobiology; human; molecular biophysics; mouse; nucleotide sugar; sialic acid; structural biology; transporter.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biological Transport
  • Crystallography, X-Ray
  • Cytidine Monophosphate / chemistry*
  • Cytidine Monophosphate / metabolism*
  • Cytidine Monophosphate N-Acetylneuraminic Acid / chemistry*
  • Cytidine Monophosphate N-Acetylneuraminic Acid / metabolism*
  • Mice
  • Protein Binding
  • Protein Conformation

Substances

  • Cytidine Monophosphate N-Acetylneuraminic Acid
  • Cytidine Monophosphate

Associated data

  • PDB/6OH3
  • PDB/6OH4