scholarly journals A neutral endopeptidase in the microvillar membrane of pig intestine. Partial purification and properties

1980 ◽  
Vol 191 (2) ◽  
pp. 645-648 ◽  
Author(s):  
E M Danielsen ◽  
J P Vyas ◽  
A J Kenny
Keyword(s):  
Brush Border ◽  
Chelating Agents ◽  
Neutral Endopeptidase ◽  
Partial Purification ◽  
Immunological Properties ◽  
Neutral Proteinase ◽  
Purification And Properties ◽  
Intestinal Enzyme ◽  
Triton X

An enzyme hydrolysing [125I]iodo-insulin B chain was enriched in preparations of intestinal microvilli. The activity could be solubilized by Triton X-100 and was partially (76-fold) purified. It was very sensitive to inhibition by phosphoramidon and was also inhibited by chelating agents. In its enzymic, molecular and immunological properties the intestinal enzyme closely resembled kidney microvillar neutral endopeptidase (kidney-brush-border neutral proteinase, EC 3.4.24.11).

scholarly journals Characterization of neutral endopeptidase 24.11 in dog glomeruli

1993 ◽  
Vol 291 (3) ◽  
pp. 773-779 ◽  
Author(s):  
C Landry ◽  
P Santagata ◽  
W Bawab ◽  
M C Fournié-Zaluski ◽  
B P Roques ◽  
...  
Keyword(s):  
Cell Surface ◽  
Brush Border ◽  
Neutral Endopeptidase ◽  
Glycosylation Pattern ◽  
Electrophoretic Mobilities ◽  
Mammalian Tissues ◽  
Dog Kidney ◽  
A Cell ◽  
Membrane Bound ◽  
Triton X

Neutral endopeptidase (NEP; also known as neprilysin and enkephalinase; EC 3.4.24.11) is a cell-surface metallopeptidase that is present in many mammalian tissues. It is particularly abundant on the brush-border membranes of the kidney proximal tubule. In this paper, the presence of NEP in purified glomeruli from dog kidney was assessed by measuring phosphoramidon- and thiorphan-sensitive [D-Ala2,Leu5]enkephalin-degrading activity. Using this assay, the Km and kcat. of the glomerular enzyme were found to be identical to those of the tubular enzyme. By Western blotting the apparent M(r) of the glomerular enzyme was found to be 104,000, compared with 94,000 for the tubular enzyme. This might be due to a different glycosylation pattern, since endoglycosidase F treatment of NEP obtained from both tissues yielded deglycosylated enzymes with similar electrophoretic mobilities. The glomerular enzyme also appears to be membrane-bound, since it was retained in the detergent-rich phase after phase separation with Triton X-114. Autoradiography experiments performed with RB104, a new highly selective and potent NEP inhibitor, showed that NEP was expressed in both glomeruli and proximal tubules. The presence in glomeruli of NEP and some other brush-border peptidases (dipeptidyl-dipeptidase IV, aminopeptidase N and angiotensin I-converting enzyme) suggests that cell-surface peptidases might play an important role as regulators of plasma-derived peptides in this part of the nephron.

scholarly journals Solubilization, partial purification and properties of N-methylglutamate dehydrogenase from Pseudomonas aminovorans

1977 ◽  
Vol 161 (2) ◽  
pp. 357-370 ◽  
Author(s):  
C W Bamforth ◽  
P J Large
Keyword(s):  
Cytochrome C ◽  
Electron Acceptor ◽  
Specific Activity ◽  
Partial Purification ◽  
Ph Optimum ◽  
Transport Chain ◽  
Purification And Properties ◽  
Solubilized Enzyme ◽  
Triton X ◽  
Sepharose 4B

1. Extracts of amine-grown Pseudomonas aminovorans contained a particle-bound N-methylglutamate dehydrogenase (EC 1.5.99.5). The enzyme was not present in succinate-grown cells, and activity appeared before growth began in succinate-grown cells which had been transferred to methylamine growth medium. 2. Membrane-containing preparations from methylamine-grown cells catalysed an N-methylglutamate-dependent uptake of O2 or reduction of cytochrome c, which was sensitive to inhibitors of the electron-transport chain. 3. N-Methylglutamate dehydrogenase activity with phenazine methosulphate or 2,6-dichlorophenol-indophenol as electron acceptor could be solubilized with 1% (w/v) Triton X-100. The solubilized enzyme was much less active with cytochrome c as electron acceptor and did not sediment in 1 h at 150000g. Solubilization was accompanied by a change in the pH optimum for activity. 4. The solubilized enzyme was partially purified by Sepharose 4B and hydroxyapatite chromatograpy to yield a preparation 22-fold increased in specific activity over the crude extract. 5. The partially-purified enzyme was active with sarcosine, N-methylalanine and N-methylaspartate as well as with N-methylglutamate. Evidence suggesting activity with N-methyl D-amino acids as well as with the L-forms was obtained. 6. The enzyme was inhibited by p-chloromercuribenzoate, iodoacetamide and by both ionic and non-ionic detergents. 2-Oxoglutarate and formaldehyde were also inhibitors. 7. Kinetic analysis confirmed previous workers' observations of a group transfer (Ping Pong) mechanism. 8. Spectral observations suggested that the partially purified preparation contained flavoprotein and a b-type cytochrome. 9. The role of the enzyme in the oxidation of methylamine is discussed.

Partial purification and properties of an endo-xylanase from cucumber seeds

1991 ◽  
Vol 81 (3) ◽  
pp. 327-334 ◽  
Author(s):  
Cesar V. Mujer ◽  
Dale W. Kretchman ◽  
A. Raymond Miller
Keyword(s):  
Partial Purification ◽  
Purification And Properties

Partial purification and properties of a 36-kDa 1-aminocyclopropane-l-carboxylate N-malonyltransferase from mung bean

1995 ◽  
Vol 94 (4) ◽  
pp. 629-634 ◽  
Author(s):  
Mohamed Benichou ◽  
Gracia Martinez-Reina ◽  
Felix Romojaro ◽  
Jean-Claude Pech ◽  
Alain Latche
Keyword(s):  
Mung Bean ◽  
Partial Purification ◽  
Purification And Properties

scholarly journals Partial purification and properties of a pre-mRNA splicing activity.

1985 ◽  
Vol 260 (2) ◽  
pp. 1096-1102
Author(s):  
P R DiMaria ◽  
G Kaltwasser ◽  
C J Goldenberg
Keyword(s):  
Mrna Splicing ◽  
Partial Purification ◽  
Purification And Properties

Partial Purification and Properties of Human Brain Aldehyde Dehydrogenases

1985 ◽  
Vol 45 (6) ◽  
pp. 1903-1910 ◽  
Author(s):  
Jacques-Andre Maring ◽  
Richard A. Deitrich ◽  
Roger Little
Keyword(s):  
Human Brain ◽  
Partial Purification ◽  
Aldehyde Dehydrogenases ◽  
Purification And Properties

Partial purification of a boar sperm membrane protein inducing sperm agglutinating antibody

1990 ◽  
Vol 10 (2) ◽  
pp. 131-139
Author(s):  
Oyewole Adeyemo ◽  
E. O. Okegbile ◽  
O. O. Olorunsogo
Keyword(s):  
Molecular Weight ◽  
Membrane Protein ◽  
Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Partial Purification ◽  
Boar Sperm ◽  
Sperm Membrane ◽  
Triton X

For the development of immunological contraception, attention is being concentrated on the possibility of using a sperm membrane antigen. Boar sperm membrane was extracted with triton-X 100 and fractionated by Sephadex G-150 column chromatography. The glycosylated and nonglycosylated portions of protein peaks from the gel filtration were obtained by fractionating on concanavalin A-Sepharose and eluting the bound protein with 0.3 M methyl mannoside. A glycosylated fraction was found to induce sperm agglutinating antibodies in rabbit. The partially purified protein has a molecular weight of 30 kilodaltons, as determined by sodium dodecyl polyaccyrlamide gel electrophoresis. Further work is planned on the histochemical determination of the origin of this protein and species cross-activity of the antibody.

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