Analysis of homologous and heterologous interactions between UDP-galactose transporter and beta-1,4-galactosyltransferase 1 using NanoBiT

Anal Biochem. 2020 Mar 15:593:113599. doi: 10.1016/j.ab.2020.113599. Epub 2020 Jan 28.

Abstract

Split luciferase complementation assay is one of the approaches enabling identification and analysis of protein-protein interactions in vivo. The NanoBiT technology is the most recent improvement of this strategy. Nucleotide sugar transporters and glycosyltransferases of the Golgi apparatus are the key players in glycosylation. Here we demonstrate the applicability of the NanoBiT system for studying homooligomerization of these proteins. We also report and discuss a novel heterologous interaction between UDP-galactose transporter and beta-1,4-galactosyltransferase 1.

Keywords: Beta-1,4-galactosyltransferase 1; Golgi apparatus; N-Glycosylation; NanoBiT; Split luciferase complementation assay; UDP-Galactose transporter.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Biological Transport
  • CHO Cells
  • Cricetulus
  • Golgi Apparatus / metabolism
  • HEK293 Cells
  • Humans
  • Luminescent Measurements / methods*
  • Monosaccharide Transport Proteins / metabolism*
  • N-Acetyllactosamine Synthase / metabolism*
  • Nanotechnology / methods*
  • Protein Binding

Substances

  • Monosaccharide Transport Proteins
  • UDP-galactose translocator
  • N-Acetyllactosamine Synthase