A rat kidney neutral peptidase that degrades B chain of insulin, glucagon, and ACTH: Purification by affinity chromatography and some properties

Varandani, Partab T.; Shroyer, Lois A.

doi:10.1016/0003-9861(77)90486-6

Cited by 33 publications

(4 citation statements)

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“…(2) The metallo-endopep- Vol. 199 tidase from rat kidney that degrades the insulin B-chain, purified by Varandani & Shroyer (1977) has weak or no activity towards proteins (denatured haemoglobin, albumin, insulin, ribonuclease) and a pH optimum of 6.5. This enzyme is reportedly inhibited by phosphate (Phelps et al, 1979).…”

Section: Discussionmentioning

confidence: 99%

“…of 93 000 (Kerr & Kenny, 1974b), the rat kidney endopeptidase has a mol.wt. of 213000 (Varandani & Shroyer, 1977), the rat parathyrin-hydrolysing enzyme has a mol.wt. of 800000 (Maruyama et al, 1970), the monkey peptidase a mol.wt.…”

Section: Discussionmentioning

confidence: 99%

“…general proteolytic activity (Hackenthal et al, 1978); kallikrein (EC 3.4.21.8) liberates bradykinin and kallidin from precursors, but has little general proteolytic activity (Pisano, 1975); and a 'postproline cleaving enzyme' has been shown to be a serine endopeptidase with limited activity towards large proteins (Koida & Walter, 1976). In addition, four metallo-endopeptidases from kidney have been described, all of which have limited or no activity towards large proteins such as haemoglobin, albumin or casein: (1) Kerr & Kenny (1974a,b) isolated a neutral metallo-endopeptidase (EC 3.4.24.11) from rabbit kidney brush border that hydrolyses insulin B chain and glucagon; (2) Varandani & Shroyer (1977) purified a membrane-bound metallo-endopeptidase from rat that hydrolyses insulin B chain;…”

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confidence: 99%

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Purification and characterization of a metallo-endoproteinase from mouse kidney

Beynon¹,

Shannon²,

Bond³

1981

Biochemical Journal

139

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A metallo-endoproteinase was purified from mouse kidney. The enzyme was solubilized from the 100 000 g sediment of kidney homogenates with toluene and trypsin, and further purified by fractionation with (NH4)2SO4. DEAE-cellulose chromatography and gel filtration. The molecular weight of the metalloproteinase was estimated by gel filtration on Sepharose 6B to be 270 000--320 000. On sodium dodecyl sulphate/polyacrylamide-gel electrophoresis in the presence of 2-mercaptoethanol, a single major protein with a mol.wt. of 81 000 was observed. Thus the active enzyme is an oligomer, probably a tetramer. It is a glycoprotein and has an apparent isoelectric point of 4.3. Kidney homogenates and purified preparations of the metalloproteinase degraded azocasein optimally at pH 9.5 and at I 0.15--0.2. The activity was not affected by inhibitors of serine proteinases (di-isopropyl phosphorofluoridate, phenylmethanesulphonyl fluoride), cysteine proteinases (4-hydroxymercuribenzoate, iodoacetate), aspartic proteinases (pepstatin) or several other proteinase inhibitors from actinomycetes (leupeptin, antipain and phosphoramidon). Inhibition of the enzyme was observed with metal chelators (EDTA, EGTA, 1,10-phenanthroline), and thiol compounds (cysteine, glutathione, dithioerythritol, 2-mercaptoethanol). The metalloproteinase degraded azocasein, azocoll, casein, haemoglobulin and aldolase, but showed little or no activity against the synthetic substrates benzoylarginine 2-naphthylamide, benzoylglycylarginine, benzyloxycarbonylglutamyltyrosine or acetylphenylalanyl 2-naphthyl ester. This metalloproteinase from mouse kidney appears to be distinct from previously described kidney proteinases.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

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Purification and characterization of a metallo-endoproteinase from mouse kidney

Beynon¹,

Shannon²,

Bond³

1981

Biochemical Journal

139

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“…Of mammalian tissues the kidney is a major source of metallo-endopeptidase activity (Erdos & Yang, 1967;Aswanikumar & Radhakrishnan, 1975;Varandani & Shroyer, 1977;Sogawa et al, 1981). One of the best characterized of these enzymes is the rabbit kidney brush-border neutral endopeptidase (NEP; EC 3.4.24.11) described by Kenny and co-workers (Kerr & Kenny, 1974a,b;Kenny, 1977); this enzyme is optimally active at neutral pH values, is sensitive to inhibition by phosphoramidon (the thermolysin inhibitor found in Streptomyces spp.…”

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confidence: 99%

Certain mouse strains are deficient in a kidney brush-border metallo-endopeptidase activity

Bond¹,

Shannon²,

Beynon³

1983

Biochemical Journal

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Preparations of microvilli from kidneys of BALB/c mice contain an alkaline metallo-endopeptidase, meprin (metallo-endopeptidase from renal tissue). Certain genealogically related inbred mice are markedly deficient in meprin activity. The meprin-deficient strains (CBA/J and C3H/HeJ) exhibit normal levels of other brush-border enzymes: alkaline phosphatase, aminopeptidase M and another proteinase, a phosphoramidon-sensitive neutral endopeptidase. Meprin deficiency cannot be attributed to a shift in pH optimum and is unlikely to be due to the presence of endogenous inhibitors.

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