Intra- and intermolecular interactions in sucrose transporters at the plasma membrane detected by the split-ubiquitin system and functional assays

Anke Reinders, Waltraud Schulze, Safia Thaminy, Igor Stagljar, Wolf B. Frommer, John M. Ward

Research output: Contribution to journalArticle

48 Citations (Scopus)

Abstract

Interaction of two separately expressed halves of sucrose transporter SUT1 was detected by an optimized split-ubiquitin system. The halves reconstitute sucrose transport activity at the plasma membrane with affinities similar to the intact protein. The halves do not function independently, and an intact central loop is not required for membrane insertion, plasma membrane targeting, and transport. Under native conditions, the halves associate into higher molecular mass complexes. Furthermore, the N-terminal half of the low-affinity SUT2 interacts functionally with the C-terminal half of SUT1. Since the N terminus of SUT2 determines affinity for sucrose, the reconstituted chimera has lower affinity than SUT1. The split-ubiquitin system efficiently detects intramolecular interactions in membrane proteins, and can be used to dissect transporter structure.

Original languageEnglish (US)
Pages (from-to)763-772
Number of pages10
JournalStructure
Volume10
Issue number6
DOIs
StatePublished - Jan 1 2002

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Ubiquitin
Sucrose
Cell Membrane
Membrane Proteins
Membranes
Proteins

Keywords

  • Assembly
  • Dimer
  • Heteromeric interaction
  • Membrane protein
  • Structure
  • Sucrose transporter

Cite this

Intra- and intermolecular interactions in sucrose transporters at the plasma membrane detected by the split-ubiquitin system and functional assays. / Reinders, Anke; Schulze, Waltraud; Thaminy, Safia; Stagljar, Igor; Frommer, Wolf B.; Ward, John M.

In: Structure, Vol. 10, No. 6, 01.01.2002, p. 763-772.

Research output: Contribution to journalArticle

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