Molecular cloning of the alpha-subunit of rat endopeptidase-24.18 (endopeptidase-2) and co-localization with endopeptidase-24.11 in rat kidney by in situ hybridization.
Endopeptidase-24.18 (endopeptidase-2, EC 3.4.24.18, E-24.18) is a Zn-ectoenzyme of rat renal and intestinal microvillar membranes exhibiting an oligomeric structure, alpha 2-beta 2. The primary structure of the alpha-subunit of E-24.18 has been defined by molecular cloning and its expression mapped in rat kidney by in situ hybridization. A 2.9-kb cDNA coding for the alpha-subunit was isolated and sequenced. It had an open reading frame of 2,244 base pairs coding for a type I membrane protein of 748 amino acids. The deduced amino acid sequence showed 87% identity with that of meprin A, a mouse metallo-endopeptidase, sharing common properties with the rat enzyme, and 85% identity with the human intestinal enzyme, 'PABA-peptide hydrolase'. Northern blot analysis revealed the alpha-subunit to be encoded by a single mRNA species of 3.2-kb. In situ hybridization performed on rat kidney showed a co-localization of E-24.18 with endopeptidase-24.11 in proximal tubules of juxtamedullary nephrons, suggesting that the two enzymes have similar or complementary physiological functions in kidney.
Corbeil
D
D
Département de biochimie, Faculté de médicine, Université de Montréal, Canada.
Gaudoux
F
F
Wainwright
S
S
Ingram
J
J
Kenny
A J
AJ
Boileau
G
G
Crine
P
P
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D003001:Cloning, Molecular; D007668:Kidney / Q000201:enzymology*; D008666:Metalloendopeptidases / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D015260:Neprilysin / Q000235:genetics*; D009693:Nucleic Acid Hybridization; D051381:Rats
D008666:Metalloendopeptidases; D015260:Neprilysin; C053833:meprin A