Identification and Characterisation of a pH-stable GFP.
Green fluorescent proteins (GFPs) are invaluable tools for modern cell biology. Even though many properties of GFP have been successfully engineered, a GFP retaining brightness at low pH has not emerged. This limits the use of GFP in quantitative studies performed in fluctuating or acidic conditions. We report the engineering and characterisation of tandem dimer GFP (pH-tdGFP), a bright and stable GFP that can be efficiently excited and maintains its fluorescence properties in acidic conditions. Therefore, pH-tdGFP could act as a quantitative marker for cellular processes that occur at low pH, such as endocytosis, autophagy or starvation.
ETH Zurich D-BSSE, Mattenstr. 26, 4058 Basel, Switzerland.
Tania Michelle
TM
Roberts
ETH Zurich D-BSSE, Mattenstr. 26, 4058 Basel, Switzerland.|Swiss Institute of Bioinformatics, Mattenstr. 26, 4058 Basel, Switzerland.
Fabian
F
Rudolf
ETH Zurich D-BSSE, Mattenstr. 26, 4058 Basel, Switzerland.
Andreas
A
Meyer
ETH Zurich D-BSSE, Mattenstr. 26, 4058 Basel, Switzerland.
Rene
R
Pellaux
ETH Zurich D-BSSE, Mattenstr. 26, 4058 Basel, Switzerland.|Swiss Institute of Bioinformatics, Mattenstr. 26, 4058 Basel, Switzerland.
Ellis
E
Whitehead
ETH Zurich D-BSSE, Mattenstr. 26, 4058 Basel, Switzerland.
Sven
S
Panke
ETH Zurich D-BSSE, Mattenstr. 26, 4058 Basel, Switzerland.
Martin
M
Held
D003952:Diagnostic Imaging / Q000379:methods*; D019281:Dimerization; D004705:Endocytosis; D004926:Escherichia coli / Q000235:genetics; D005453:Fluorescence; D049452:Green Fluorescent Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D006863:Hydrogen-Ion Concentration; D011487:Protein Conformation; D015202:Protein Engineering; D055550:Protein Stability; D012440:Saccharomyces / Q000235:genetics
D049452:Green Fluorescent Proteins