Biosynthesis and characterization of the brain-specific membrane protein DPPX, a dipeptidyl peptidase IV-related protein.
Dipeptidyl peptidase IV-related protein (DPPX) was found to be preferentially expressed in the brain tissue. We isolated two rat cDNA clones encoding DPPX-S and DPPX-L from a brain cDNA library, of which DPPX-L had a longer sequence at the NH2 terminus. The biosynthesis of DPPXs was examined in both in vitro and in vivo systems. In the cell-free translation system, DPPX-S and DPPX-L were synthesized as 93-kDa and 97-kDa forms, respectively, which are in good agreement with the molecular masses estimated from their primary structure. In COS-1 cells transfected with the cDNAs, DPPX-S and DPPX-L were initially synthesized as 113-kDa and 117-kDa forms, respectively, with high-mannose type oligosaccharides, which were then converted to 115-kDa and 120-kDa forms, mostly with the complex-type sugar chains. Immunofluorescence-microscopic observations confirmed that both DPPXs were expressed on the cell surface. DPPXs were found to have no enzyme activity of DPPIV, even when they were mutated to have the consensus active-site sequence Gly-X-Ser-X-Gly for serine proteases. Immunoblot analysis of samples prepared from various rat tissues demonstrated that DPPX-S, but not DPPX-L, was detectable only in the brain tissue. These results indicate that, of the two isoforms, DPPX-S is preferentially expressed in the brain tissue as the surface glycoprotein without protease activity, although its function remains unknown at present.
Kin
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Department of Biochemistry, Fukuoka University School of Medicine, Nanakuma, Jonan-ku, Fukuoka 814-0180, Japan.
Misumi
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Y
Ikehara
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Y
D000818:Animals; D001921:Brain / Q000201:enzymology / Q000378:metabolism*; D019556:COS Cells / Q000201:enzymology / Q000378:metabolism; D003001:Cloning, Molecular / Q000379:methods; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D018819:Dipeptidyl Peptidase 4 / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D006023:Glycoproteins / Q000378:metabolism*; D066298:In Vitro Techniques; D008565:Membrane Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D009928:Organ Specificity; D014176:Protein Biosynthesis; D051381:Rats; D012697:Serine Endopeptidases; D014158:Transcription, Genetic
D018076:DNA, Complementary; D006023:Glycoproteins; D008565:Membrane Proteins; D018819:Dipeptidyl Peptidase 4; D012697:Serine Endopeptidases