Superfolder GFP Is Fluorescent in Oxidizing Environments When Targeted via the Sec Translocon

Deborah E. Aronson, Lindsey M. Costantini, Erik L. Snapp

Research output: Contribution to journalArticle

73 Citations (Scopus)

Abstract

The ability to study proteins in live cells using genetically encoded fluorescent proteins (FPs) has revolutionized cell biology Researchers have created numerous FP biosensors and optimized FPs for specific organisms and subcellular environments in a rainbow of colors However, expressing FPs in oxidizing environments such as the eukaryotic endoplasmic reticulum (ER) or the bacterial periplasm can impair folding, thereby preventing fluorescence A substantial fraction of enhanced green fluorescent protein (EGFP) oligomerizes to form non-fluorescent mixed disulfides in the ER and EGFP does not fluoresce in the periplasm when targeted via the SecYEG translocon To overcome these obstacles, we exploited the highly efficient folding capability of superfolder GFP (sfGFP) Here, we report sfGFP does not form disulfide-linked oligomers in the ER and maltose-binding protein (MBP) signal sequence (peri)-sfGFP is brightly fluorescent in the periplasm of Escherichia coli. Thus, sfGFP represents an important research tool for studying resident proteins of oxidizing environments.

Original languageEnglish (US)
Pages (from-to)543-548
Number of pages6
JournalTraffic
Volume12
Issue number5
DOIs
StatePublished - May 2011

Fingerprint

Periplasm
Endoplasmic Reticulum
Proteins
Disulfides
Cytology
Maltose-Binding Proteins
Biosensing Techniques
Protein Sorting Signals
Oligomers
Biosensors
Escherichia coli
Cell Biology
Color
Fluorescence
Research Personnel
Research
enhanced green fluorescent protein

Keywords

  • ER
  • Maltose-binding protein
  • Periplasm
  • Superfolder GFP
  • Translocon

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Structural Biology
  • Molecular Biology
  • Genetics

Cite this

Superfolder GFP Is Fluorescent in Oxidizing Environments When Targeted via the Sec Translocon. / Aronson, Deborah E.; Costantini, Lindsey M.; Snapp, Erik L.

In: Traffic, Vol. 12, No. 5, 05.2011, p. 543-548.

Research output: Contribution to journalArticle

Aronson, Deborah E. ; Costantini, Lindsey M. ; Snapp, Erik L. / Superfolder GFP Is Fluorescent in Oxidizing Environments When Targeted via the Sec Translocon. In: Traffic. 2011 ; Vol. 12, No. 5. pp. 543-548.
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