Molecular cloning of the α‐subunit of rat endopeptidase‐24.18 (endopeptidase‐2) and co‐localization with endopeptidase‐24.11 in rat kidney by in situ hybridization

FEBS Letters

Gaudoux, Florence¹  Boileau, Guy¹  Wainwright, Sandra¹  Crine, Philippe¹  Kenny, A.John²  Corbeil, Denis¹  Ingram, Jean² 

[1] Département de biochimie, Faculté de médicine, Université de Montréal, Montréal, Canada;Membrane Peptidase Research Group, Department of Biochemistry and Molecular Biology, University of Leeds, Leeds, LS2 9JT, UK

关键词: Membrane protein;    Endopeptidase-2;    cDNA;    Rat kidney;    In situ hybridization;    E-24.18;    endopeptidase-2;    E-24.11;    endopeptidase-24.11;    PPH;    N-benzoyl-l-tyrosyl-p-aminobenzoic acid hydrolase;    bp;    base pair;    kb;    kilobase;    SDS;    sodium dodecyl sulphate;    PAGE;    polyacrylamide gel eletrophoresis;   

DOI  :  10.1016/0014-5793(92)81095-4

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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 oligometric structure, α₂-β₂. The primary structure of the α-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 α-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 α-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.

【 授权许可】

Unknown