RESUMO
Meprin-a is a metalloendopeptidase present at high levels in the kidney brush border of some inbred mouse strains. Meprin-b is a latent metallo-endopeptidase, activated by trypsin-mediated proteolysis in vitro, that is present at similar activities (after activation) in all mouse strains. Meprin (a mixture of a and b forms) was purified from a high-meprin Mep-1a/a animal, and Lys-C peptides of this preparation were sequenced. The sequence data were used to direct the synthesis of peptides that were conjugated to albumin and used as immunogens. One of these antisera was specific to meprin-b and thus provided a specific tool to monitor expression of this form of meprin in different mouse strains.
Assuntos
Tiopronina/imunologia , Sequência de Aminoácidos , Animais , Western Blotting , Hidrólise , Rim/enzimologia , Camundongos , Camundongos Endogâmicos , Microvilosidades/enzimologia , Dados de Sequência Molecular , Papaína , Mapeamento de Peptídeos , Especificidade da Espécie , Tiopronina/química , TripsinaRESUMO
The brush border membrane of mice and rats contains a phosphoramidon-insensitive metalloproteinase, meprin (neutral endopeptidase-2; NEP-2). The role of meprin is unknown, but we have shown that urine from these species contains insulin B chain degrading activity that is due to a phosphoramidon-insensitive metalloendopeptidase. By enzymic and immunological criteria, it is likely that this activity is due to meprin, and introduces the possibility that this enzyme may have a role in urinary function.
Assuntos
Metaloendopeptidases/urina , Animais , Rim/enzimologia , Camundongos , Microvilosidades/enzimologia , Ratos , Ratos EndogâmicosRESUMO
1. Inbred mouse strains differ markedly in the expression of a kidney brush border metalloendopeptidase, meprin-a. 2. Brush border preparations from mice of the low-meprin-a phenotype (specific activities less than 5% of the high-meprin-a trait) contain a metallo-endopeptidase, meprin-b, that is larger than meprin-a, and which is inactive unless the membrane preparations are treated with trypsin. 3. This cryptic metallo-endopeptidase has been previously postulated to be a stalled precursor of meprin-a. 4. We show here that meprin-b is present in all mice-high and low meprin-a phenotypes--and that this activity is similar in substrate specificity and amount present in the brush border. 5. Meprin-b may therefore be a distinct gene product that is independent of meprin-a phenotype.
