Chemosensing ensembles for monitoring biomembrane transport in real time.
The efficacy of drugs and biomolecules relies on their ability to pass through the bilayer. The development of methods to directly and sensitively monitor these membrane transport processes has remained an experimental challenge. A macrocyclic host (p-sulfonatocalix[4]arene or cucurbit[7]uril) and a fluorescent dye (lucigenin or berberine) are encapsulated as a chemosensing ensemble inside liposomes, which allows for a direct, real-time fluorescence monitoring of the passage of unlabeled bioorganic analytes. This in vitro assay is transferable to different channel proteins and analytes, has potential for fluorescence-based screening, e.g., of channel modulators, and yields the absolute kinetics of translocation. Using this new biophysical method, we observed for the first time direct rapid translocation of protamine, an antimicrobial peptide, through the bacterial transmembrane protein OmpF.
School of Engineering and Science, Jacobs University Bremen, Campus Ring 1, 28759 Bremen (Germany).
Garima
G
Ghale
Adrienne G
AG
Lanctôt
Hannah T
HT
Kreissl
Maik H
MH
Jacob
Helge
H
Weingart
Mathias
M
Winterhalter
Werner M
WM
Nau
D000166:Acridines / Q000737:chemistry*; D001599:Berberine / Q000737:chemistry*; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000737:chemistry*; D008081:Liposomes / Q000737:chemistry* / Q000378:metabolism; D008958:Models, Molecular; D015394:Molecular Structure; D011479:Protamines / Q000737:chemistry / Q000378:metabolism*; D013997:Time Factors
D000166:Acridines; D005456:Fluorescent Dyes; D008081:Liposomes; D011479:Protamines; D001599:Berberine; C033472:10,10'-dimethyl-9,9'-biacridinium
antimicrobial peptides; calixarenes; fluorescence; macrocycles; membrane proteins