Calcium-activated neutral protease (CANP) in normal and dysmyelinating mutant paralytic tremor rabbit myelin.
Calcium-activated neutral protease (CANP) in normal and dysmyelinating mutant, paralytic tremor (PT) rabbit myelin and premyelin fractions was studied using immature (4-5 wk) or adult animals. The enzyme was estimated by determination of its catalytic activity as well as by using immunoblot analysis after SDS-PAGE separation. The presence of two forms of CANP--one activated by calcium in the micromolar concentration (mu CANP) range and the other exhibiting low calcium sensitivity in the millimolar concentration range (m-CANP)--was found in the myelin and premyelin fractions. The developmental pattern of the enzyme activity was different for each of these two enzyme isoforms depending on the fraction studied. The higher activity on CANP (both isoforms) found in PT myelin and premyelin could be related to delayed myelination and/or to the higher turnover rate of already formed myelin. These results suggest complex and specific roles for these isoenzymes during myelin formation as is discussed further in this article. Our results confirm the extensive degradation of myelin basic protein (MBP), proteolipid protein (PLP), and, to a lesser extent, the other myelin proteins by endo- and exogenous CANP. This degradation process was significantly elevated in PT rabbit myelin. Moreover as was shown by two-dimensional gel electrophoresis, calcium-controlled proteolysis in nonmutant rabbits affected the net-charge of MBP in a manner similar to that reported for PT myelin, suggesting the possible involvement of CANP in the generation of charge isomers of MBP.
Domańska-Janik
K
K
Department of Neurochemistry, Polish Academy of Sciences, Warsaw.
de Nechaud
B
B
Inomata
M
M
Kawashima
S
S
Zalewska
T
T
D000818:Animals; D001923:Brain Chemistry; D002154:Calpain / Q000032:analysis*; D003711:Demyelinating Diseases / Q000235:genetics / Q000378:metabolism*; D007527:Isoenzymes / Q000032:analysis*; D009124:Muscle Proteins / Q000032:analysis*; D009154:Mutation; D009185:Myelin Proteins / Q000378:metabolism; D009186:Myelin Sheath / Q000201:enzymology*; D010243:Paralysis / Q000201:enzymology / Q000235:genetics; D011817:Rabbits / Q000235:genetics / Q000378:metabolism*; D014202:Tremor / Q000201:enzymology / Q000235:genetics
D007527:Isoenzymes; D009124:Muscle Proteins; D009185:Myelin Proteins; D002154:Calpain