SYNTHESIS, CHARACTERIZATION AND FLUORESCENCE STUDIES ON N‐α‐DANSYLALANYLLYSYL TRYPSINO (HIS‐46)‐METHANE
- 12 January 1979
- journal article
- research article
- Published by Wiley in International Journal of Peptide and Protein Research
- Vol. 13 (1) , 95-101
- https://doi.org/10.1111/j.1399-3011.1979.tb01854.x
Abstract
The 1-dimethylamino-5-naphthalene sulfonyl (dansyl) derivative of alanyllsyl chloromethane was synthesized and employed to introduce the fluorescent dansyl moiety specifically into the active site of trypsin via affinity labelling. The potential of dansylalanyllysyl chloromethane lies in its high degree of selectivity and markedly faster rate of enzyme inactivation when compared to previously synthesized, single residue affinity label chromophores. This permits the practical utilization of stoichiometric amounts of the inhibitor to achieve 100% inactivation of trypsin, even at high dilutions. The transfer of energy between the four tryptophan residues of trypsin and the bound dansyl group has been investigated in the fluorescent inhibitor-enzyme conjugate. From transfer efficiency measurements mean distances of 19.0 Å and 19.3 Å between the point of attachment of the dansyl group and the four tryptophan residues of trypsin have been calculated. These compare well with the mean value of 18.8 Å derived from calculations based on crystallo-graphic model studies.Keywords
This publication has 23 references indexed in Scilit:
- Fluorescence study of hybrid hemoglobins containing free base and zinc protoporphyrin IXBiochemistry, 1974
- Peptide chloromethyl ketones as irreversible inhibitors of elastaseBiochemistry, 1973
- SPECIFIC FLUORESCENT DERIVATIVES OF MACROMOLECULES.International Journal of Peptide and Protein Research, 1972
- Structure of crystalline α-chymotrypsinJournal of Molecular Biology, 1972
- Uses of Fluorescence in Biophysics: Some Recent DevelopmentsAnnual Review of Biophysics and Bioengineering, 1972
- Design and synthesis of inhibitors for crystallographic studies on the active site of chymotrypsinJournal of the American Chemical Society, 1970
- Intramolecular singlet excitation transfer. Applications to polypeptidesBiochemistry, 1969
- Nonradiative Energy Transfer in Systems in which Rotatory Brownian Motion is FrozenThe Journal of Chemical Physics, 1968
- Identification of the histidine residue at the active center of trypsin labelled by TLCKBiochemical and Biophysical Research Communications, 1967
- Zur Spezifität von Chymotrypsin und TrypsinHoppe-Seyler´s Zeitschrift Für Physiologische Chemie, 1966