Development of membrane-based biosensors: Measurement of current from photocycling bacteriorhodopsin on patch clamp electrodes

Journal Article · Fri Jan 01 00:00:00 EST 1988 · Adv. Exp. Med. Biol.; (United States)

DOI:https://doi.org/10.1007/978-1-4684-7908-9_20· OSTI ID:5769124

Yager, P

Our initial work toward developing membrane protein-based biosensors has involved use of bacteriorhodopsin (BR) as a model membrane protein. BR was incorporated into liposomes of a polymerizable lecithin, and was shown to pump protons in response to illumination both before and after polymerization of the lipids. In the work described in this paper, BR was first reincorporated in liposomes of asolectin by consonication with purple membrane. The liposomes, which sustained the function of the protein, were used to form a monolayer at the air-water interface. This monolayer was transferred as a bilayer onto patch electrode. When illuminated with a pulse of 514.5-nm light the lipid/protein patch produced a current spike into the pipette corresponding to events no later than the generation of the 412-nm intermediate, probably caused by pumping of protons across the patch membrane. The experiment demonstrates not only the extreme sensitivity of amperometric detection, but also a small tendency for membrane proteins to preferentially orient in this configuration.

• Research Organization: Naval Research Laboratory, Washington, DC (USA)
• OSTI ID: 5769124
• Journal Information: Adv. Exp. Med. Biol.; (United States), Vol. 238
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
PROTONS
MEMBRANE TRANSPORT
RHODOPSIN
BIOELECTRICITY
LIPIDS
LIPOSOMES
PHOSPHOLIPIDS
PHOTOCHEMISTRY
PHOTOSYNTHETIC MEMBRANES
POLYMERIZATION
BARYONS
CELL CONSTITUENTS
CHEMICAL REACTIONS
CHEMISTRY
ELECTRICITY
ELEMENTARY PARTICLES
ESTERS
FERMIONS
HADRONS
MEMBRANES
NUCLEONS
ORGANIC COMPOUNDS
ORGANIC PHOSPHORUS COMPOUNDS
ORGANOIDS
PIGMENTS
PROTEINS
550200* - Biochemistry