Organization of membrane proteins in the intact myelin sheath. Pyridoxal phosphate and salicylaldehyde as probes of myelin structure

Abstract

Pyridoxal phosphate and salicylaldehyde were used as protein-labeling probes to study the organization of membrane proteins in the intact myelin sheath of the cat dorsal column. Both reagents react with protein amino groups to form Schiff's bases which can be reduced with NaB3H4. The relatively membrane-impermeant pyridoxal phosphate labels all proteins of the intact myelin except basic protein. This major protein of myelin is labeled only after loss of membrane integrity. The relatively membrane-permeant probe, salicylaldehyde, was then used to establish that the basic protein is truly located on the cytoplasmic side of the myelin bilayer, and not merely sequestered within the multiple lamellar structure of the sheath. All proteins in the intact myelin are readily labeled by this reagent, with the label distribution pattern identical to that of disrupted myelin fragments. These data suggest a model for myelin structure in which the basic protein is the only major protein component located exclusively on the cytoplasmic side of the membrane (the major period zone of the sheath), with the other major proteins disposed wholly, or in part, in the extracellular half of the membrane bilayer (the intraperiod zone). All proteins, although asymmetrically disposed with respect to membrane sidedness, appear to be randomly distributed throughout the lamellae which comprise the sheath.