Loose interaction between glyceraldehyde-3-phosphate dehydrogenase and phosphoglycerate kinase revealed by fluorescence resonance energy transfer-fluorescence lifetime imaging microscopy in living cells

Yosuke Tomokuni, Kenji Goryo, Ayako Katsura, Satoru Torii, Ken Ichi Yasumoto, Klaus Kemnitz, Mamiko Takada, Hiroshi Fukumura, Kazuhiro Sogawa

Research output: Contribution to journalArticlepeer-review

10 Citations (Scopus)

Abstract

Loose interaction between the glycolytic enzymes glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and phosphoglycerate kinase (PGK) was visualized in living CHO-K1 cells by fluorescence resonance energy transfer (FRET), using time-domain fluorescence lifetime imaging microscopy. FRET between active tetrameric subunits of GAPDH linked to cerulean or citrine was observed, and this FRET signal was significantly attenuated by coexpression of PGK. Also, direct interaction between GAPDH-citrine and PGK-cerulean was observed by FRET. The strength of FRET signals between them was dependent on linkers that connect GAPDH to citrine and PGK to cerulean. A coimmunoprecipitation assay using hemagglutinin-tagged GAPDH and FLAG-tagged PGK coexpressed in CHO-K1 cells supported the FRET observation. Taken together, these results demonstrate that a complex of GAPDH and PGK is formed in the cytoplasm of living cells.

Original languageEnglish
Pages (from-to)1310-1318
Number of pages9
JournalFEBS Journal
Volume277
Issue number5
DOIs
Publication statusPublished - 2010 Mar

Keywords

  • FLIM
  • FRET
  • GAPDH
  • Loose interaction
  • PGK

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Fingerprint Dive into the research topics of 'Loose interaction between glyceraldehyde-3-phosphate dehydrogenase and phosphoglycerate kinase revealed by fluorescence resonance energy transfer-fluorescence lifetime imaging microscopy in living cells'. Together they form a unique fingerprint.

Cite this