The increased ecto-5'-nucleotidase activity in muscle, heart and liver of laminin alpha2-deficient mice is not caused by an elevation in the mRNA content.
We have previously shown that mouse muscle and liver contain catalytically active and inactive ecto-5'-nucleotidase (eNT) variants and that eNT activity in these tissues increases in laminin alpha2 (merosin)-deficient Lama2dy mice. These results prompted us to study whether: (1) the increase of eNT activity depends on the change in the content of merosin between healthy and dystrophic organs; (2) the active and inactive eNT variants arise from the same or distinct mRNAs; (3) the enhancement of the activity is caused by an increase in the eNT mRNA content. Compared to healthy organs, the activity in dystrophic organs increased four-fold in muscle, 1.7-fold in liver, 1.4-fold in heart and not at all in kidney and lung. The level of immunolabelled eNT protein per unit of activity suggested a similar ratio of inactive to active eNT in healthy liver, kidney, heart and muscle, which increased greatly in lung. The size of the eNT subunit in liver, kidney, heart and muscle (72 kDa) decreased to 66 kDa in lung. The identification of a single RT-PCR product suggested that active and inactive eNT arise from the same mRNA and are generated by a differential post-translational processing. Compared to the content in muscle, the amount of eNT mRNA was 12-fold higher in liver and kidney, eight-fold in heart and five-fold in lung. The relative content of eNT mRNA was unaffected by the deficiency of merosin.
Morote-García
Julio C
JC
Departamento de Bioquímica y Biología Molecular-A, Edificio de Veterinaria, Universidad de Murcia, Apdo. 4021, E-30071 Espinardo, Murcia, Spain.
Sánchez Del Campo
Luis F
LF
Campoy
Francisco J
FJ
Vidal
Cecilio J
CJ
Muñoz-Delgado
Encarnación
E
D015720:5'-Nucleotidase / Q000032:analysis / Q000378:metabolism*; D000818:Animals; D004789:Enzyme Activation / Q000235:genetics*; D007797:Laminin / Q000737:chemistry / Q000172:deficiency* / Q000235:genetics; D008099:Liver / Q000737:chemistry / Q000378:metabolism*; D051379:Mice; D008817:Mice, Mutant Strains / Q000235:genetics; D018482:Muscle, Skeletal / Q000737:chemistry / Q000378:metabolism*; D009137:Muscular Dystrophy, Animal / Q000235:genetics / Q000378:metabolism; D009206:Myocardium / Q000737:chemistry / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism*
D007797:Laminin; D012333:RNA, Messenger; C102645:laminin alpha 2; D015720:5'-Nucleotidase