Temperature-pressure stability of green fluorescent protein: A Fourier transform infrared spectroscopy study

Carsten H. Scheyhing, Filip Meersman, Matthias A. Ehrmann, Karel Heremans, Rudi F. Vogel

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44 Scopus citations


Green fluorescent protein (GFP) is widely used as a marker in molecular and cell biology. For its use in high-pressure microbiology experiments, its fluorescence under pressure was recently investigated. Changes in fluorescence with pressure were found. To find out whether these are related to structural changes, we investigated the pressure stability of wild-type GFP (wtGFP) and three of its red shift mutants (AFP, GFPmut1, and GFPmut2) using Fourier transform infrared spectroscopy. For the wt GFP, GFPmut1, and GFPmut2 we found that up to 13-14 kbar the secondary structure remains intact, whereas AFP starts unfolding around 10 kbar. The 3-D structure is held responsible for this high-pressure stability. Previously observed changes in fluorescence at low pressure are rationalized in terms of the pressure-induced elastic effect. Above 6 kbar, loss of fluorescence is due to aggregation. Revisiting the temperature stability of GFP, we found that an intermediate state is populated along the unfolding pathway of wtGFP. At higher temperatures, the unfolding resulted in the formation of aggregates of wtGFP and its mutants.

Original languageEnglish
Pages (from-to)244-253
Number of pages10
Issue number4
StatePublished - 15 Nov 2002
Externally publishedYes


  • FTIR spectroscopy
  • Green fluorescent protein
  • Red shift mutants
  • Temperature-pressure stability


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