TY - JOUR
T1 - Interactions of fusidic acid and elongation factor G with lipid membranes
AU - Muhonen, Jaana
AU - Vidgren, Jukka
AU - Helle, Anne
AU - Yohannes, Gebrenegus
AU - Viitala, Tapani
AU - Holopainen, Juha M.
AU - Wiedmer, Susanne K.
N1 - Funding Information:
Financial support was received from the Academy of Finland (SA 202216, 114292) (SKW) and Sigrid Juselius Foundation, The Helsinki University Central Hospital Research Fund, and Evald and Hilda Nissi Foundation (JMH). The authors thank Dr. Jari Metso (Department of Molecular Medicine, National Public Health Institute, Helsinki) for help with the radioactivity measurements. The donation of EF-G by Dr. Suparna Sanyal (Department of Cell and Molecular Biology, Uppsala University, Sweden) is much appreciated.
PY - 2008/3/1
Y1 - 2008/3/1
N2 - Fusidic acid (FA) is a potent antibiotic and blocks the protein synthesis by binding to elongation factor G (EF-G) directly. Here we hypothesized that the antibiotic activity of FA would be potentiated by several orders of magnitude if both FA and EF-G would be residing in the lipid membranes and, hence, the probability of interaction would transform from three-dimensional to two-dimensional. Such detailed information could lead to more effective therapeutic interventions if they are understood on a molecular level. Interactions between FA and various lipid membranes composed of 1-palmitoyl-2-oleyl-sn-glycero-3-phosphocholine (POPC) and cholesterol (Chol) were studied by capillary electrochromatography (CEC). The influence of the lipid vesicle size-sonicated liposomes and liposomes extruded through 30-, 50-, and 100-nm filters-on the packing of vesicles on the silica capillary surface was investigated by CEC and dissipative quartz crystal microbalance. The CEC results evidenced that FA interacts with and resides in phospholipid membranes. Likewise, monolayer, asymmetrical flow field flow fractionation, and CEC studies confirmed that EF-G is hydrophobic and incorporated into POPC and POPC/Chol membranes. Including EF-G in phospholipid vesicles did not improve the binding of FA to the membranes.
AB - Fusidic acid (FA) is a potent antibiotic and blocks the protein synthesis by binding to elongation factor G (EF-G) directly. Here we hypothesized that the antibiotic activity of FA would be potentiated by several orders of magnitude if both FA and EF-G would be residing in the lipid membranes and, hence, the probability of interaction would transform from three-dimensional to two-dimensional. Such detailed information could lead to more effective therapeutic interventions if they are understood on a molecular level. Interactions between FA and various lipid membranes composed of 1-palmitoyl-2-oleyl-sn-glycero-3-phosphocholine (POPC) and cholesterol (Chol) were studied by capillary electrochromatography (CEC). The influence of the lipid vesicle size-sonicated liposomes and liposomes extruded through 30-, 50-, and 100-nm filters-on the packing of vesicles on the silica capillary surface was investigated by CEC and dissipative quartz crystal microbalance. The CEC results evidenced that FA interacts with and resides in phospholipid membranes. Likewise, monolayer, asymmetrical flow field flow fractionation, and CEC studies confirmed that EF-G is hydrophobic and incorporated into POPC and POPC/Chol membranes. Including EF-G in phospholipid vesicles did not improve the binding of FA to the membranes.
KW - Capillary electromigration techniques
KW - Elongation factor G
KW - Field flow fractionation
KW - Fusidic acid
KW - Liposomes
KW - Quartz crystal microbalance
UR - http://www.scopus.com/inward/record.url?scp=38649137725&partnerID=8YFLogxK
U2 - 10.1016/j.ab.2007.10.014
DO - 10.1016/j.ab.2007.10.014
M3 - Article
C2 - 17980694
AN - SCOPUS:38649137725
SN - 0003-2697
VL - 374
SP - 133
EP - 142
JO - Analytical Biochemistry
JF - Analytical Biochemistry
IS - 1
ER -