Cleavage of the P0 glycoprotein of the rat peripheral nerve myelin: tentative identification of cleavage site and evidence for the precursor-product relationship.
The incubation of sciatic nerve slices in Krebs Ringer bicarbonate (KRB) buffer (pH 7.4) at 37 degrees C, or the incubation of freshly isolated myelin in ammonium bicarbonate buffer (pH 8), resulted in the generation of a 24 kDa protein with a concomitant decrease of P0 protein. The conversion of P0 into 24 kDa protein was blocked by heating isolated myelin at 100 degrees C for 5 min suggesting that the reaction is enzyme mediated. Inclusion of the protease inhibitors and chelating agent to isolated myelin did not prevent the formation of 24 kDa protein. Similarly, addition of CaCl2 to isolated myelin did not accentuate the formation of 24 kDa protein suggesting that the conversion of P0 into 24 kDa protein may not be due to Ca2+ activated protease. It is postulated that the formation of 24 kDa protein may be due to neutral protease and/or metalloproteinase associated with the PNS myelin. 24 kDa protein was purified and characterized. The N-terminal sequence of 1-17 amino acid residues of 24 kDa protein was identical to P0. 24 kDa protein was immunostained and immunoprecipitated with anti-P0 antiserum indicating the immunological similarities between P0 and 24 kDa protein. Labeling of 24 kDa protein with [35S]methionine provided evidence that P0 may be in all probability cleaved between Met-168 and Met-193. Further studies were carried out to demonstrate that 24 kDa protein was phosphorylated, glycosylated and acylated like P0. Phosphorylation of 24 kDa protein in the nerve slices was increased five-fold by phorbol esters and phosphoserine was the only phosphoamino acid identified after partial acid hydrolysis of 24 kDa protein.(ABSTRACT TRUNCATED AT 250 WORDS)
Agrawal
H C
HC
Department of Pediatrics, Washington University School of Medicine, St. Louis, Missouri 63110.
Agrawal
D
D
Strauss
A W
AW
D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel; D066298:In Vitro Techniques; D007700:Kinetics; D008562:Membrane Glycoproteins / Q000378:metabolism; D008970:Molecular Weight; D018993:Myelin P0 Protein; D009185:Myelin Proteins / Q000302:isolation & purification / Q000378:metabolism*; D009186:Myelin Sheath / Q000378:metabolism*; D010766:Phosphorylation; D051381:Rats; D012584:Sciatic Nerve / Q000378:metabolism*
D008562:Membrane Glycoproteins; D018993:Myelin P0 Protein; D009185:Myelin Proteins