Direct AFM observation of saposin C-induced membrane domains in lipid bilayers: from simple to complex lipid mixtures.
Saposin C (Sap C) is a small glycoprotein required by glucosylceramidase (GCase) for hydrolysis of glucosylceramide to ceramide and glucose in lysosomes. The molecular mechanism underlying Sap C stimulation of the enzyme activation is not fully understood. Here, atomic force microscopy (AFM) has been used to study Sap C-membrane interactions under physiological conditions. First, to establish how Sap C-membrane interactions affect membrane structure, lipid bilayers containing zwitterionic and anionic phospholipids were used. It was observed that Sap C induced two types of membrane restructuring effects, i.e., the formation of patch-like domains and membrane destabilization. Bilayers underwent extensive structural reorganization. To validate the biological importance of the membrane restructuring effects, interaction of Sap C with lipid bilayers composed of cholesterol, sphingomyelin, and zwitterionic and anionic phospholipids were studied. Although similar membrane restructuring effects were observed, Sap C-membrane interactions, in this case, were remarkably modulated and their effects were restricted to a limited area. As a result, nanometer-sized domains were formed. The establishment of a model membrane system will allow us to further study the dynamics, structure and mechanism of the Sap C-associated membrane domains and to examine the important role that these domains may play in enzyme activation.
You
Hong Xing
HX
Department of Cell Biology, Neurobiology, and Anatomy, College of Medicine, University of Cincinnati, 3125 Eden Ave, Cincinnati, OH 45267-0521, USA. hong.you@uc.edu
Qi
Xiaoyang
X
Yu
Lei
L
D045424:Complex Mixtures / Q000737:chemistry; D008051:Lipid Bilayers / Q000737:chemistry*; D008560:Membrane Fluidity; D021962:Membrane Microdomains / Q000737:chemistry* / Q000648:ultrastructure*; D008565:Membrane Proteins / Q000737:chemistry; D018625:Microscopy, Atomic Force; D010743:Phospholipids / Q000737:chemistry*; D049231:Saposins / Q000737:chemistry* / Q000648:ultrastructure*; D013499:Surface Properties
D045424:Complex Mixtures; D008051:Lipid Bilayers; D008565:Membrane Proteins; D010743:Phospholipids; D049231:Saposins