Acquiring snapshots of the orientation of trans-membrane protein domains using a hybrid FRET pair.
FEBS Lett
; 589(8): 885-9, 2015 Apr 02.
Article
in En
| MEDLINE
| ID: mdl-25747388
ABSTRACT
One challenge in studying the function of membrane-embedded proteins is determining the orientation of key domains in the context of the changing and dynamic membrane environment. We describe a confocal microscopy setup that utilizes external electric field pulses to direct dipicrylamine (DPA) to a membrane leaflet. The detection of FRET between DPA and a fluorescent probe attributes it to the inner or outer leaflet of a membrane. By utilizing short acquisition times and confocal imaging, this attribution could be made even in changing membrane environments. Our setup adds versatility to the study of the biological activity of membrane-embedded proteins.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Fluorescence Resonance Energy Transfer
/
Membrane Proteins
Limits:
Animals
Language:
En
Journal:
FEBS Lett
Year:
2015
Type:
Article
Affiliation country:
United States