Real-Time Imaging of Protease Action on Substrates Covalently Immobilised to Polymer Supports.
Adv Synth Catal
; 349(8-9): 1321-1326, 2007 Jun.
Article
en En
| MEDLINE
| ID: mdl-19779571
ABSTRACT
We report for the first time single bead spatially resolved activity measurements of solid-phase biocatalytic systems followed in real-time. Trypsin cleavage of Bz-Arg-OH and subtilisin cleavage of Z-Gly-Gly-Leu-OH each liberate a free amino group on aminocoumarin covalently immobilised to PEGA(1900) beads [a co-polymer of poly(ethylene glycol) with molecular mass of 1900 cross-linked with acrylamide]. This restores fluorescence which is imaged in optical sections by two-photon microscopy. For trypsin cleavage, fluorescence is restricted initially to surface regions, with more than 1 hour needed before reaction is fully underway in the bead centre, presumably reflecting slow enzyme diffusion. In contrast, for subtilisin cleavage fluorescence develops throughout the bead more quickly.
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1
Colección:
01-internacional
Base de datos:
MEDLINE
Idioma:
En
Revista:
Adv Synth Catal
Año:
2007
Tipo del documento:
Article
País de afiliación:
Reino Unido