Effect of cholesterol content on affinity and stability of factor VIII and annexin V binding to a liposomal bilayer membrane.
To investigate the effect of cholesterol composition on the binding of factor VIII (FVIII) and annexin V (AV) to membranes, liposomal membranes with phospholipid bilayers of various compositions of phosphatidylcholine (PC), phosphatidylserine (PS), and cholesterol were constructed. A surface plasmon resonance (SPR) biosensor system was employed to measure the equilibrium and rate constants of the bindings. As expected, PS was found to play a dominant role in the binding of AV; its binding level was directly proportional to the PS composition in a liposome. The binding levels of FVIII and AV to liposome increased with an increase in cholesterol composition in liposome. It seemed to suggest that cholesterol in liposome acts as a 'phospholipid arrangement' factor by inducing the formation of PS-rich microdomains. However, in the absence of PS (20% on a mole basis), cholesterol could not exert the binding enhancement effect, which again confirmed the critical role of PS in the bindings. Stability of the AV binding was significantly improved by the increase in cholesterol content; for AV, the dissociation rate constant was decreased approximately fivefold, from 1.7 x 10(-3)s(-1) in the absence of cholesterol to 3.3 x 10(-4)s(-1) in the presence of only 10% cholesterol. But, for FVIII the binding stability was not so much influenced by the cholesterol addition (up to 50% on a mole basis). In summary, by using liposomes on an SPR system, we were able to demonstrate quantitatively the apparent effects of cholesterol on the binding affinity and stability of the membrane-binding proteins.
Department of Bionanotechnology, Hanyang University, Ansan, Republic of Korea.
Jun Yeoung
JY
Jeon
Sang Youn
SY
Hwang
Seung Hun
SH
Cho
Jaebum
J
Choo
E K
EK
Lee
D017304:Annexin A5 / Q000737:chemistry*; D002784:Cholesterol / Q000737:chemistry*; D005169:Factor VIII / Q000737:chemistry*; D008051:Lipid Bilayers / Q000737:chemistry*; D008081:Liposomes / Q000737:chemistry*; D011485:Protein Binding
D017304:Annexin A5; D008051:Lipid Bilayers; D008081:Liposomes; D005169:Factor VIII; D002784:Cholesterol