title: Meprin A, the major matrix degrading enzyme in renal tubules, produces a novel nidogen fragment in vitro and in vivo.
abstract: We examined the effect of meprin A, the major matrix degrading metalloproteinase in rat kidney, on the laminin-nidogen complex. N-terminal sequence information from the most abundant 55 kDa fragment revealed that it was a breakdown product of nidogen rather than laminin. In comparison with over 50 nidogen cleavage sites produced by other proteases, the meprin A-induced nidogen cleavage site at amino acid position 899-900, a glutamine-glycine site in the G3 domain, is unique. In addition, these data demonstrate that meprin A degrades the G3 domain of nidogen even in the presence of laminin binding, which usually accords protection from proteolytic degradation. Meprin A also degraded purified nidogen into similar breakdown products. Given that the tubular basement membrane is located on the basilar side of the cell, the location of meprin A on the apical brush border makes it difficult to envision a role for meprin A in injury-induced basement membrane component breakdown. Thus, we examined the possibility that following renal tubular epithelial cell injury, meprin A undergoes a translocation to reach the underlying basement membrane. After renal ischemia-reperfusion there was a marked alteration in meprin A staining with meprin A now distributed throughout the renal tubular cell cytoplasm and directly adherent to the tubular basement membrane. This was in contrast to the usual linear staining of the brush border of tubules in the corticomedullary junction. These data provide unequivocal evidence that following injury, meprin A undergoes redistribution and/or adherence to the tubular basement membrane. Since in our in vitro studies, we identified a distinct meprin-induced 55 kDa nidogen breakdown product, the urine was also examined for the presence of nidogen degradation products after rat renal ischemia-reperfusion injury. Western blots showed a marked increase in the urinary 55 kDa nidogen fragment as early as the first day following ischemia-reperfusion injury and continuing for six days. Taken together, these in vivo data strongly support the notion that the nidogen breakdown products are the result of partial degradation of tubular basement membrane by meprin A following renal tubular ischemia-reperfusion injury.
authors.affiliation: Department of Pathology, University of Arkansas for Medical Sciences, Little Rock, USA. pdwalker@life.uams.edu
authors.forename: P D
authors.initials: PD
authors.lastname: Walker
authors.forename: G P
authors.initials: GP
authors.lastname: Kaushal
authors.forename: S V
authors.initials: SV
authors.lastname: Shah
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D005109:Extracellular Matrix / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D007511:Ischemia / Q000378:metabolism; D007672:Kidney Cortex / Q000378:metabolism; D007684:Kidney Tubules / Q000201:enzymology*; D007797:Laminin / Q000378:metabolism; D008297:Male; D008562:Membrane Glycoproteins / Q000096:biosynthesis* / Q000235:genetics; D008666:Metalloendopeptidases / Q000378:metabolism*; D010446:Peptide Fragments / Q000096:biosynthesis* / Q000235:genetics; D051381:Rats; D012079:Renal Circulation / Q000502:physiology; D015427:Reperfusion Injury / Q000378:metabolism; D014018:Tissue Distribution
chemical_list: D007797:Laminin; D008562:Membrane Glycoproteins; D010446:Peptide Fragments; C029967:nidogen; D008666:Metalloendopeptidases; C053833:meprin A