title: Tryptophan mediated photoreduction of disulfide bond causes unusual fluorescence behaviour of Fusarium solani pisi cutinase.
abstract: The fluorescence signal of the single tryptophan residue (Trp69) of Fusarium solani pisi cutinase is highly quenched. However, prolonged irradiation of the enzyme in the tryptophan absorption band causes an increase of the tryptophan fluorescence quantum yield by an order of magnitude. By using a combination of NMR spectroscopy and chemical detection of free thiol groups with a sulfhydryl reagent we could unambiguously show that the unusual fluorescence behaviour of Trp69 in cutinase is caused by the breaking of the disulfide bond between Cys31 and Cys109 upon irradiation, while the amide-aromatic hydrogen bond between Ala32 and Trp69 remains intact. This is the first example of tryptophan mediated photoreduction of a disulfide bond in proteins.
authors.affiliation: NSR Center for Molecular Structure, Design and Synthesis, Laboratory of Biophysical Chemistry, University of Nijmegen, The Netherlands.
authors.forename: J J
authors.initials: JJ
authors.lastname: Prompers
authors.forename: C W
authors.initials: CW
authors.lastname: Hilbers
authors.forename: H A
authors.initials: HA
authors.lastname: Pepermans
mesh_terms: D000409:Alanine / Q000737:chemistry; D000577:Amides / Q000737:chemistry; D002265:Carboxylic Ester Hydrolases / Q000737:chemistry* / Q000528:radiation effects*; D003545:Cysteine / Q000737:chemistry; D004220:Disulfides / Q000737:chemistry; D005453:Fluorescence; D005670:Fusarium / Q000201:enzymology*; D006860:Hydrogen Bonding; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D009587:Nitrogen Isotopes; D013438:Sulfhydryl Compounds / Q000737:chemistry; D013439:Sulfhydryl Reagents / Q000737:chemistry; D014364:Tryptophan / Q000737:chemistry / Q000528:radiation effects*
chemical_list: D000577:Amides; D004220:Disulfides; D009587:Nitrogen Isotopes; D013438:Sulfhydryl Compounds; D013439:Sulfhydryl Reagents; D014364:Tryptophan; D002265:Carboxylic Ester Hydrolases; C019952:cutinase; D003545:Cysteine; D000409:Alanine