A novel metalloproteinase present in freshly isolated rat glomeruli

Q. Le, S. Shah, Justin H Nguyen, S. Cortez, W. Baricos

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

We have utilized [3H]gelatin to document high activity of a metalloproteinase present in freshly isolated rat glomeruli. [3H]gelatin degradation by glomeruli was markedly inhibited by EDTA (10 mM: -89 ± 2.3%) and o-phenanthroline (2 mM: -72 ± 0.1%), inhibitors of metalloproteinases. No significant inhibition of [3H]gelatin degradation was observed with inhibitors of serine or cysteine proteinases. Most (>80%) of the glomerular metalloproteinase (GLOMP) activity was associated with the pellet after centrifugation of sonicated glomeruli at 100,000 g for 90 min. The pH optimum for gelatin degradation by sonicated glomeruli was ~pH 8.5. Sodium dodecyl sulfate substrate (gelatin)-polyacrylamide gel electrophoresis revealed a single major band of EDTA-inhibitable gelatin-degrading activity with a molecular mass of ~116-125 kDa. The GLOMP activity was not inhibited by tissue inhibitors of metalloproteinases, did not appear to be latent, and was not activated by organomercurial activators of several latent metalloproteinases. GLOMP activity was increased 3.4-fold after incubation with trypsin (20 μg/ml, 25 min, 22°C). These data indicate that GLOMP is distinct from the previously described matrix metalloproteinases, as well as other metalloproteinases present in the kidney, including the gelatinase secreted by cultured mesangial cells, Meprin, and endopeptidase 24.11 (enkephalinase, EC 3.4.24.11).

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Renal Fluid and Electrolyte Physiology
Volume260
Issue number4 29/4
StatePublished - 1991
Externally publishedYes

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Metalloproteases
Gelatin
Neprilysin
Edetic Acid
Tiopronin
Cysteine Proteinase Inhibitors
Tissue Inhibitor of Metalloproteinases
Gelatinases
Serine Proteinase Inhibitors
Mesangial Cells
Matrix Metalloproteinases
Centrifugation
Sodium Dodecyl Sulfate
Trypsin
Polyacrylamide Gel Electrophoresis
Cultured Cells
Kidney

Keywords

  • Kidney
  • Proteinase

ASJC Scopus subject areas

  • Physiology

Cite this

A novel metalloproteinase present in freshly isolated rat glomeruli. / Le, Q.; Shah, S.; Nguyen, Justin H; Cortez, S.; Baricos, W.

In: American Journal of Physiology - Renal Fluid and Electrolyte Physiology, Vol. 260, No. 4 29/4, 1991.

Research output: Contribution to journalArticle

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