Lysosomal heterogeneity of dipeptidyl peptidase II active on collagen-related peptides.
The subcellular distribution of dipeptidyl peptidase II (DPP II) in the rat kidney cortex, as determined by subfractionation of the mitochondrial/lysosomal fraction by rate sedimentation, indicated that this enzyme is mainly associated with the large, fast sedimenting lysosomes (protein droplets). The small lysosomes, on the other hand, displayed considerable size heterogeneity as indicated by the broad distribution of DPP II; cathepsin B, and a tripeptidyl peptidase active on Gly-Pro-Met-2-naphthylamide at pH 4 (TPP 4). Cathepsin D and N-acetyl-beta-D-glucosaminidase were limited primarily to the slower-sedimenting, small lysosomes. Equilibrium banding in sucrose gradients of the two main DPP II-containing lysosomal populations showed that the large lysosomes banded at a density of 1.235-1.24 g/ml while small lysosomes banded at three densities: 1.11-1.15 g/ml (lysosomal fragments), 1.20 g/ml (light lysosomes), and 1.235 g/ml (dense lysosomes). Identical distribution pattern were obtained for DPP II using either Lys-Ala-7-(4-methyl)coumarylamide or Gly-Pro-2-naphthylamide as the substrate at pH 5.5 and 5.0, respectively. Notably, DPP II and TPP 4, and cathepsin B as well, gave banding densities and distributions that were consistent with a lysosomal localization. Since triplets of the Gly-Pro-X-type released by the TPP 4 are ideal substrates for DPP II, the integrated action of tripeptidyl and dipeptidyl peptidases could make a novel contribution to the renal depolymerization and reabsorption of polypeptides, in particular the proline-rich, collagen-derived sequences that possess repeating-triplet primary structures.
Andersen
K J
KJ
University of Bergen, Medical Department A, Haukeland Sykehus, Bergen, Norway.
McDonald
J K
JK
D000626:Aminopeptidases; D000818:Animals; D002499:Centrifugation, Density Gradient; D003094:Collagen / Q000378:metabolism*; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases / Q000378:metabolism*; D066298:In Vitro Techniques; D007672:Kidney Cortex / Q000201:enzymology* / Q000648:ultrastructure; D008247:Lysosomes / Q000201:enzymology*; D008297:Male; D010455:Peptides / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains; D012697:Serine Endopeptidases / Q000378:metabolism
D010455:Peptides; D003094:Collagen; D000626:Aminopeptidases; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases; C048001:tripeptidyl-peptidase 2; C100151:dipeptidyl peptidase II; D012697:Serine Endopeptidases