Characterization of the interaction between phospholipid and protein by capillary electrophoresis with laser-induced fluorescence detection.
We report an electrophoretic mobility shift-based method to study the interactions between phospholipids and proteins by capillary electrophoresis with laser-induced fluorescence detection. A fluorogenic dye, 3-(2-furoyl)quinoline-2-carboxaldehyde (FQ), was used to label phosphatidylserine (PS). Then the FQ labeled PS (FQ-PS) was used as the fluorescent probe for monitoring the association between PS and bovine serum albumin (BSA). Two conjugates were observed to form between each PS species and BSA, indicating that two interactions exist between these PS species and BSA. We can also detect the competitive association with BSA between labeled PS and unlabeled PS. This method only needs a minute volume of sample. It is highly sensitive and can be used to detect the interaction between phospholipids and nanomolar concentrations of proteins, such as BSA.
Hu
S
S
Department of Chemistry, University of Washington, Seattle 98195-1700, USA.
Zhang
L
L
Dovichi
N J
NJ
D019075:Electrophoresis, Capillary / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry; D005663:Furans / Q000737:chemistry; D007834:Lasers; D010718:Phosphatidylserines / Q000378:metabolism*; D011804:Quinolines / Q000737:chemistry; D012680:Sensitivity and Specificity; D012710:Serum Albumin, Bovine / Q000378:metabolism*; D013050:Spectrometry, Fluorescence / Q000379:methods*
D005456:Fluorescent Dyes; D005663:Furans; D010718:Phosphatidylserines; D011804:Quinolines; C063756:3-(2-furoyl)quinoline-2-carbaldehyde; D012710:Serum Albumin, Bovine