Multimeric Structure of the Secreted Meprin A Metalloproteinase and Characterization of the Functional Protomer

Faoud Ishmael, Mona T. Norcum, Stephen Benkovic, Judith S. Bond

Research output: Contribution to journalArticle

40 Citations (Scopus)

Abstract

Meprin A secreted from kidney and intestinal epithelial cells is capable of cleaving growth factors, extracellular matrix proteins, and biologically active peptides. The secreted form of meprin A is a homo-oligomer composed of α subunits, a multidomain protease of 582 amino acids coded for near the major histocompatibility complex of the mouse and human genome. Analyses of the recombinant homo-oligomeric form of mouse meprin A by gel filtration, nondenaturing gel electrophoresis, and cross-linking (with disuccinimidyl suberate or N-(4-azido-2,3,5,6-tetraflourobenzyl)-3-maleimidylpropionamide) indicate that the secreted enzyme forms high molecular weight multimers, with a predominance of decamers. The multimers are composed of disulfide-linked dimers attached noncovalently by interactions involving the meprin, A5 protein, receptor protein-tyrosine phosphatase μ (MAM) domain. The active protomer is the noncovalently linked dimer. Linkage of active protomers by disulfide-bonds results in an oligomer of ∼900 kDa, which is unique among proteases and distinguishes meprin A as the largest known secreted protease. Electron microscopy revealed that the protein was present in two states, a crescent-shaped structure and a closed ring. It is concluded from this and other data that the covalent attachment of the protomers enables noncovalent associations of the native enzyme to form higher oligomers that are critical for hydrolysis of protein substrates.

Original languageEnglish (US)
Pages (from-to)23207-23211
Number of pages5
JournalJournal of Biological Chemistry
Volume276
Issue number25
DOIs
StatePublished - Jun 22 2001

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meprin A
Protein Subunits
Metalloproteases
Oligomers
Peptide Hydrolases
Disulfides
Dimers
Gels
Tiopronin
Proteins
Protein Tyrosine Phosphatases
Extracellular Matrix Proteins
Human Genome
Enzymes
Major Histocompatibility Complex
Electrophoresis
Electron microscopy
Gel Chromatography
Hydrolysis
Intercellular Signaling Peptides and Proteins

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Ishmael, Faoud ; Norcum, Mona T. ; Benkovic, Stephen ; Bond, Judith S. / Multimeric Structure of the Secreted Meprin A Metalloproteinase and Characterization of the Functional Protomer. In: Journal of Biological Chemistry. 2001 ; Vol. 276, No. 25. pp. 23207-23211.
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Multimeric Structure of the Secreted Meprin A Metalloproteinase and Characterization of the Functional Protomer. / Ishmael, Faoud; Norcum, Mona T.; Benkovic, Stephen; Bond, Judith S.

In: Journal of Biological Chemistry, Vol. 276, No. 25, 22.06.2001, p. 23207-23211.

Research output: Contribution to journalArticle

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