Identification of inactive ecto-5'-nucleotidase in normal mouse muscle and its increased activity in dystrophic Lama2(dy) mice.
Ecto-5'-nucleotidase (eNT) activity and protein in normal (NM) and merosin-deficient dystrophic (DM) Lama2(dy) mice muscle were studied. eNT activity in DM was three- to four-fold that in NM. eNT in NM and DM displayed the same kinetic properties. Slot and Western blotting revealed that the immunoreactive protein was two to three times more abundant in control muscle, when NM and DM samples with the same eNT activity were compared, indicating that mouse muscle contains catalytically inactive eNT components. eNT activity and protein peaks coincided in sedimentation analyses, revealing that inactive eNT occurs as dimers. Most eNT activity and protein of NM bound to Lens culinaris (LCA) or Ricinus communis (RCA) agglutinins, but half of the activity and one-third of the protein bound to wheat germ agglutinin (WGA). Although WGA interaction did not permit full separation of inactive eNT, the results suggest that similar proportions of active species with and without WGA reactivity occur in mouse muscle, whereas a great fraction of the inactive eNT variants lack WGA reactivity. Because the level of eNT protein was little modified in DM, the higher eNT activity in dystrophic than in control muscle may result from misregulation in the synthesis of active and inactive eNT species or from conversion of inactive into active components.
García-Ayllón
M S
MS
Departamento de Bioquímica y Biología Molecular-A, Universidad de Murcia, Espinardo, Murcia, Spain.
Campoy
F J
FJ
Vidal
C J
CJ
Muñoz-Delgado
E
E
D015720:5'-Nucleotidase / Q000378:metabolism*; D000249:Adenosine Monophosphate / Q000378:metabolism; D000818:Animals; D057927:Hydrophobic and Hydrophilic Interactions; D007700:Kinetics; D007797:Laminin / Q000172:deficiency* / Q000235:genetics; D037102:Lectins / Q000737:chemistry; D051379:Mice; D008817:Mice, Mutant Strains; D018482:Muscle, Skeletal / Q000201:enzymology* / Q000503:physiopathology; D009137:Muscular Dystrophy, Animal / Q000201:enzymology* / Q000235:genetics / Q000503:physiopathology; D013347:Subcellular Fractions; D013501:Surface-Active Agents / Q000378:metabolism
D007797:Laminin; D037102:Lectins; D013501:Surface-Active Agents; C102645:laminin alpha 2; D000249:Adenosine Monophosphate; D015720:5'-Nucleotidase