Abstract
We labeled hyaluronan (HA) with two fluorophores, fluorescein amine and rhodamine B amine. These two fluorophores are suitable for a fluorescence (Foerster) resonance energy transfer (FRET) which results in a fluorescein quenching and an enhanced rhodamine emission. Such labeled HA (HA-FRET) is a potential sensor for HA degradation. We studied fluorescence properties of HA-FRET in the absence and presence of hyaluronidase enzyme (HA-ase). The time-resolved fluorescence measurements indicate more than 50% of FRET in the absence of HA-ase. In the presence of HA-ase FRET decreases with time, and relative fluorescence intensities of fluorescein and rhodamine shifts to fluorescein indicating a release of FRET. The kinetics of the digestion process of HA by HA-ase depends on the concentration of the enzyme. We demonstrate that simultaneous measurements of green and red emission of HA-FRET can be used in ratio metric detection of the HA-ase presence and activity. This in turn, can be utilized for the construction of a robust but reliable HA-ase sensing device.
| Original language | English |
|---|---|
| Pages (from-to) | 473-477 |
| Number of pages | 5 |
| Journal | Journal of Photochemistry and Photobiology B: Biology |
| Volume | 104 |
| Issue number | 3 |
| DOIs | |
| State | Published - 2 Sep 2011 |
Keywords
- FRET
- Fluorescence
- Hyaluronan
- Hyaluronidase
- Ratio metric sensing