Giant proteoliposomes prepared by freezing-thawing without use of detergent: reconstitution of biomembranes usually inaccessible to patch-clamp pipette microelectrode.
When sarcoplasmic reticulum membrane vesicles or synaptosomes were mixed with sonicated phospholipid vesicles and subjected to freezing-thawing, giant vesicles of up to 50 microns in diameter were formed. When the biomembrane vesicles were labeled with a covalently binding fluorescent dye, the resultant giant vesicles were fluorescent, thereby suggesting that the freezing-thawing process induces fusion of phospholipid and biomembrane vesicles. When membranes of the giant proteoliposomes thus prepared were studied using the patch-clamp technique, potassium channels of the biomembranes were detectable. The present method of the giant proteoliposome preparation is simple and rapid, and provides a system suitable for the study of ion channels of various biomembranes usually inaccessible to a patch-pipette microelectrode.
Saito
Y
Y
Faculty of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan.
Hirashima
N
N
Kirino
Y
Y
D000818:Animals; D002458:Cell Fractionation; D002499:Centrifugation, Density Gradient; D002540:Cerebral Cortex / Q000648:ultrastructure; D005615:Freezing; D007425:Intracellular Membranes / Q000502:physiology / Q000648:ultrastructure*; D008081:Liposomes; D008297:Male; D008564:Membrane Potentials; D008839:Microelectrodes; D009132:Muscles / Q000648:ultrastructure; D010713:Phosphatidylcholines; D010743:Phospholipids; D011510:Proteolipids; D011817:Rabbits; D051381:Rats; D011919:Rats, Inbred Strains; D012519:Sarcoplasmic Reticulum / Q000502:physiology / Q000648:ultrastructure*; D013574:Synaptosomes / Q000502:physiology / Q000648:ultrastructure*
D008081:Liposomes; D010713:Phosphatidylcholines; D010743:Phospholipids; D011510:Proteolipids; C020485:proteoliposomes; C024990:asolectin