Cryoelectron microscopy of protein-lipid complexes of human myelin basic protein charge isomers differing in degree of citrullination.
Myelin basic protein (MBP) is considered to be essential for the maintenance of stability of the myelin sheath. Reduction in cationicity of MBP, especially due to conversion of positively charged arginine residues to uncharged citrulline (Cit), has been found to be associated with multiple sclerosis (MS). Here, the interactions of an anionic phosphatidylserine/monosialoganglioside-G(M1) (4:1, w:w) lipid monolayer with 18.5-kDa MBP preparations from age-matched adult humans without MS (no Cit residues), with chronic MS (6 Cit), and with acute Marburg-type MS (18 Cit) were studied by transmission and ultralow dose scanning transmission electron microscopy under cryogenic conditions. Immunogold labeling and single particle electron crystallography were used to define the nature of the complexes visualized. These electron microscopical analyses showed that the three different MBP charge isomers all formed uniformly sized and regularly shaped protein-lipid complexes with G(M1), probably as hexamers, but exhibited differential association with and organization of the lipid. The least cationic Marburg MBP-Cit(18) formed the most open protein-lipid complex. The data show a disturbance in lipid-MBP interactions at the ultrastructural level that is related to degree of citrullination, and which may be involved in myelin degeneration in multiple sclerosis.
Beniac
D R
DR
Department of Molecular Biology and Genetics, University of Guelph, Guelph, Ontario, N1G 2W1, Canada.
Wood
D D
DD
Palaniyar
N
N
Ottensmeyer
F P
FP
Moscarello
M A
MA
Harauz
G
G
D000328:Adult; D001120:Arginine / Q000737:chemistry; D001327:Autoimmune Diseases / Q000378:metabolism; D002956:Citrulline / Q000032:analysis*; D020285:Cryoelectron Microscopy; D005677:G(M1) Ganglioside / Q000737:chemistry; D006801:Humans; D007091:Image Processing, Computer-Assisted; D007150:Immunohistochemistry; D046911:Macromolecular Substances; D008855:Microscopy, Electron, Scanning; D009103:Multiple Sclerosis / Q000378:metabolism; D004676:Myelin Basic Protein / Q000737:chemistry / Q000648:ultrastructure*; D010718:Phosphatidylserines / Q000737:chemistry; D020033:Protein Isoforms / Q000737:chemistry / Q000648:ultrastructure*; D011499:Protein Processing, Post-Translational
D046911:Macromolecular Substances; D004676:Myelin Basic Protein; D010718:Phosphatidylserines; D020033:Protein Isoforms; D002956:Citrulline; D005677:G(M1) Ganglioside; D001120:Arginine