Preparation of fluorescent derivatives of lipases and their use in fluorescence energy transfer studies in hydrocarbon/water interfaces

Gareth Roberts, M. P. Tombs

Research output: Contribution to journalArticlepeer-review

Abstract

Fluorescein isothiocyanate reacted with a chromobacter and pseudomonad lipase to yield mono-substituted, fully active, enzymes. With the carbocyanine dye 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate (DiI) in the non-aqueous phase, fluorescence energy transfer was used to follow the lipase and similarly labelled model proteins in and out of the interface in heptane, and heptane/di-O-palmitoyl-rac-glycerol (a substrate analogue), emulsions. Competitive binding, and displacement by other proteins could also be followed.
Original languageEnglish
Pages (from-to)327-334
Number of pages8
JournalBBA - Biomembranes
Volume902
Issue number3
DOIs
Publication statusPublished - 3 Sep 1987

Keywords

  • fluorescence energy transfer
  • lipase
  • hydrocarbon/water interface

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