Identification of a Lysine Residue at a Nucleotide Binding Site in the Firefly Luciferase With p-fluorosulfonyl[14C]benzoyl-5'-adenosine

Biochemistry. 1981 Mar 3;20(5):1253-6. doi: 10.1021/bi00508a031.

Abstract

Firefly luciferase is 80-90% inactivated within 3 h upon incubation with the adenine nucleotide analogue p-fluorosulfonylbenzoyl-5'-adenosine (FSBA). Although 4 mol of 14C-FSBA/mol of enzyme is irreversibly bound during inactivation, only 1 mol of 14C-FSBA appears to be specifically directed to an adenine nucleotide binding site on the enzyme. The other 3 mol of 14C-FSBA is bound to the protein nonspecifically. The major radioactive peptide in a tryptic digest os labeled luciferase was isolated and shown to have the following amino acid sequence: *Lys-Gly-Glx-Asx-Ser-Lys, where *Lys is the radioactive derivative of the lysine residue that was sulfonylated during the inactivation.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Adenosine / analogs & derivatives*
  • Adenosine / pharmacology
  • Affinity Labels* / pharmacology
  • Amino Acid Sequence
  • Amino Acids / analysis
  • Animals
  • Binding Sites
  • Coleoptera / enzymology*
  • Kinetics
  • Luciferases / metabolism*
  • Lysine / analysis*
  • Peptide Fragments / analysis
  • Protein Binding
  • Trypsin

Substances

  • Affinity Labels
  • Amino Acids
  • Peptide Fragments
  • 5'-(4-fluorosulfonylbenzoyl)adenosine
  • Luciferases
  • Trypsin
  • Lysine
  • Adenosine