Brain capillary 46,000 dalton protein is cytoplasmic actin and is localized to endothelial plasma membrane.
The most abundant protein of the brain capillary, which makes up the blood-brain barrier (BBB) in vivo, is a protein that migrates at a molecular weight of approximately 46 kDa on sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The bovine brain capillary 46 kDa protein was purified by SDS-PAGE and Sephadex G-25 gel filtration. The purified protein migrated as a single band of molecular weight of approximately 42,000 Da on subsequent SDS-PAGE followed by silver staining. The protein was digested by trypsin and tryptic peptides were analyzed by reverse phase high-performance liquid chromatography (HPLC). Two of these peptides, 11 and 18 amino acids in length, were sequenced and found to be identical to amino acid sequences corresponding to portions of cytoplasmic actin. The SDS-PAGE gel-purified 46 kDa protein was also subjected to limited proteolysis using S. aureus V8 protease, and this resulted in the formation of a prominent 31 kDa doublet as well as smaller proteolytic fragments, and these fragments were of identical molecular weight to those generated from limited proteolysis of bovine actin. Electron microscopic immunoperoxidase studies with primary cultures of bovine brain capillary endothelium showed that immunoreactive actin is intimately associated with the plasma membranes. In conclusion, the brain capillary 46 kDa protein is cytoplasmic actin and is localized to the endothelial plasma membrane. Modulations of brain capillary endothelial actin may play a role in the regulation of BBB permeability.
Pardridge
W M
WM
Department of Medicine, UCLA School of Medicine 90024-1682.
Nowlin
D M
DM
Choi
T B
TB
Yang
J
J
Calaycay
J
J
Shively
J E
JE
D000199:Actins / Q000378:metabolism*; D000818:Animals; D002196:Capillaries / Q000378:metabolism; D002417:Cattle; D002462:Cell Membrane / Q000378:metabolism; D002560:Cerebrovascular Circulation; D002851:Chromatography, High Pressure Liquid; D003593:Cytoplasm / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D004730:Endothelium, Vascular / Q000378:metabolism* / Q000648:ultrastructure; D007150:Immunohistochemistry; D008970:Molecular Weight; D010449:Peptide Mapping; D011506:Proteins / Q000302:isolation & purification / Q000378:metabolism*
D000199:Actins; D011506:Proteins