Tissue distribution and subcellular localization of rabbit liver metalloendopeptidase.
We have previously isolated rabbit liver microsomal metalloendopeptidase (MEP) as a candidate for the processing enzyme of vitamin K-dependent plasma proteins. A cDNA coding for MEP has revealed that it is structurally related to metalloendopeptidase-24.15, which catalyzes the proteolytic processing of several bioactive peptides. In this study we examined the tissue distribution and subcellular localization of MEP by light and electron microscopic immunohistochemical methods, in addition to Northern blot analysis. Chicken polyclonal antibodies were raised by using synthetic peptides AG1 (Met31-Asn46) and AG3 (Asp537-Gly551) derived from the sequence of MEP. Both anti-AG1 and anti-AG3 antibodies reacted specifically with MEP, as judged by Western blotting and immunohistochemical methods. Both antibodies gave an identical staining distribution, which was localized on the luminal cell surfaces and in the cytoplasm of the following organs: liver, brain, lungs, kidneys, esophagus, stomach, duodenum, pancreas, placenta, epididymis, uterus, ovary, and oviduct. Northern blot analysis revealed that the expression of MEP mRNA is similar to its immunohistochemical distribution except in the heart. These results suggest that MEP may participate more closely in a degradation role in peptide metabolism in various tissues than in a processing role of the proprotein, like metalloendopeptidase-24.15.
Nakagawa
K
K
Department of Pathology, Faculty of Medicine, Kyushu University, Fukuoka, Japan.
Kawabata
S
S
Nakashima
Y
Y
Iwanaga
S
S
Sueishi
K
K
D000595:Amino Acid Sequence; D000818:Animals; D015152:Blotting, Northern; D001921:Brain / Q000201:enzymology; D002462:Cell Membrane / Q000201:enzymology; D003593:Cytoplasm / Q000201:enzymology; D004386:Duodenum / Q000201:enzymology; D005260:Female; D007124:Immunoenzyme Techniques; D007668:Kidney / Q000201:enzymology; D008099:Liver / Q000201:enzymology; D008168:Lung / Q000201:enzymology; D008297:Male; D008666:Metalloendopeptidases / Q000032:analysis* / Q000235:genetics; D016253:Microscopy, Immunoelectron; D008862:Microsomes, Liver / Q000201:enzymology*; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D011817:Rabbits; D013270:Stomach / Q000201:enzymology; D013737:Testis / Q000201:enzymology; D014018:Tissue Distribution
D012333:RNA, Messenger; D008666:Metalloendopeptidases