Patch clamp detection in capillary electrophoresis.
We describe a capillary electrophoresis-patch clamp (CE-PC) analysis of biomolecules that activate ligand-gated ion channels. CE-PC offers a powerful means for identifying receptor ligands based on the combination of the characteristic receptor responses they evoke and their differential electrophoretic migration rates. Corner frequencies, membrane reversal potentials, and mean and unitary single-channel receptor responses were calculated from currents recorded with patch clamp detection. This information was then combined with the electrophoretic mobility of the receptor ligand, which is proportional to the charge-to-frictional-drag ratio of that species. We applied CE-PC to separate and detect the endogenous receptor agonists gamma-aminobutyrate and L-glutamate and the synthetic glutamate receptor agonists N-methyl-D-aspartate and kainic acid. We present dose-response data for electrophoretically separated kainic acid and discuss its implications for making the CE-PC detection system quantitative.
Jardemark
K
K
Department of Anatomy and Cell Biology, Faculty of Medicine, Göteborg University, Sweden.
Orwar
O
O
Jacobson
I
I
Moscho
A
A
Zare
R N
RN
D000818:Animals; D019075:Electrophoresis, Capillary / Q000295:instrumentation / Q000379:methods*; D005260:Female; D007395:Interneurons / Q000737:chemistry / Q000378:metabolism; D015640:Ion Channel Gating / Q000502:physiology; D007473:Ion Channels / Q000737:chemistry; D008297:Male; D018408:Patch-Clamp Techniques; D051381:Rats; D017207:Rats, Sprague-Dawley
D007473:Ion Channels