Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.4.23.5 (cathepsin D)
 4,130 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Two kinds of cathepsin D were found in Japanese monkey lung and were named cathepsins D-I and D-II. Cathepsin D-I was partially purified by ammonium sulfate fractionation and DEAE-cellulose column chromatography. It had properties common to other ordinary cathepsins D in terms of the elution position from a DEAE-cellulose column at pH 8.0, the pH-dependence of activity toward acid-denatured hemoglobin, and the molecular weight of 35,000 as determined by Sephadex G-100 gel filtration. On the other hand, cathepsin D-II was purified about 1,000-fold by a combination of ammonium sulfate fractionation and column chromatographies on DEAE-cellulose and Sephadex G-100. It was a very acidic protein as judged from its elution position from a DEAE-cellulose column at pH 8.0, and the high mobility toward the anode on disc gel electrophoresis at pH 8.6. Its molecular weight was determined to be 35,000 by Sephadex G-100 gel filtration and 39,000 by SDS-polyacrylamide gel electrophoresis. It was optimally active at pH 2.8 against acid-denatured hemoglobin as a substrate, showing 80% of the optimal activity at pH 1.0, and almost no activity above pH 4.0. This pH-profile of activity was similar to that of monkey pepsin C (gastricsin). It did not hydrolyze N-acetyl-L-phenylalanyl-3,5-diiodo-L-tyrosine, a synthetic substrate for pepsin, but was inhibited by a series of pepsin inhibitors such as pepstatin, 1,2-epoxy-3-(p-nitrophenoxy)propane, p-bromophenacyl bromide, and diazoacetyl-DL-norleucine methyl ester, although the diazo reagent was a rather weak inhibitor of the enzyme. The amino acid composition of cathepsin D-II was found to be fairly different from those of other cathepsins D. However, it showed a striking resemblance to that of Japanese monkey pepsinogen C, suggesting some evolutionary relationship between them.
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PMID:The structure and function of acid proteases. VIII. Purification and characterization of cathepsins D from Japanese monkey lung. 2 23

BW-175 is a newly synthesized renin inhibitor which is a nonpeptidic, norleucine analog. Its IC50 values for renin activity in human, squirrel monkey, marmoset, dog, hog, rabbit and rat plasma were 3.3, 6.6, 2.4, 42, 110, 86 and 3500 nM, respectively, and 26 microM for cathepsin D. Pepsin and angiotensin converting enzyme were hardly inhibited at 10(-4) M. BW-175 showed an oral bioavailability of 2.8% at 10 mg/kg and 9.7% at 30 mg/kg in rats. In normotensive, furosemide-treated high-renin marmosets, BW-175 (30 mg/kg p.o.) caused an intensive reduction in plasma renin activity and plasma angiotensin I formation, associated with a reduction in systolic blood pressure of 10-20 mm Hg for 2 hours.
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PMID:A novel nonpeptidic, orally active renin inhibitor. 249 4

Cathepsin D of human leukocytes was isolated and characterized. Purified leukocytes were lysed under nitrogen pressure and the proteinase activity precipitated by centrifugation at 48,000 x g. The precipitate was extracted by various buffers. The yield of cathepsin D was almost pH-independent but could be increased by Triton X-100. Employing gel chromatography the activity was found at a molecular mass close to 42,000 Da. Purification of the enzyme was performed by a two-step procedure using pepstatin-Sepharose chromatography and ion exchange chromatography. Three multiple forms of the enzyme were separated by ion exchange chromatography. The isoelectric points of the three forms of the enzyme were close to pH 5.0. The enzyme showed the typical characteristics of the acid proteinase cathepsin D. Enzyme activity was influenced by heavy metals such as Hg2 and Fe3 as well as by typical inhibitors for carboxyl-proteinases such as diazoacetyl-DL-norleucine methyl ester, 1,2-epoxy-3-(4-nitrophenoxy)propane and 4-bromo-phenacylbromide. An immunological comparison with cathepsin D from human liver by immunodiffusion and immunoelectrophoresis indicates identity of the two enzymes.
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PMID:Cathepsin D from human leukocytes. Purification by affinity chromatography and properties of the enzyme. 314 53

A mathematical treatment for the general case of enzyme inactivation by an inhibitor that breaks down in solution in a first-order reaction is presented. Cathepsin D was inactivated by fluorescein isothiocyanate with a K(i) of 4.47mum. Kinetic constants were also determined for the inactivation of cathepsin D by 1,1-bis(diazoacetyl)-2-phenylethane, and the inactivation of pepsin C by diazoacetyl-dl-norleucine methyl ester.
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PMID:Kinetics of irreversible enzyme inhibition by an unstable inhibitor. 461 85

Procathepsin D-II (Mr = 37 500) was purified from Japanese monkey lung at pH 7.0, and was shown to be converted to the active form, cathepsin D-II (Mr = 33 000) via an intermediate (Mr = 35 500) upon treatment at pH 3.0 and 14 degrees C. Procathepsin D-II was shown to be the inactive precursor of cathepsin D-II based on the following results: the former was inactive toward heat-denaturated casein at pH 5.4 whereas the latter was active; the former was not inactivated by diazoacetyl-DL-norleucine methyl ester in the presence of Cu2+ ion at pH 6.0 whereas the latter was inactivated rapidly under the same conditions; and the former had no affinity to pepstatin-Sepharose between pH 5 and 7 whereas the latter was adsorbed to it. With a rabbit antiserum against procathepsin D-II, cathepsin D-II, pepsinogen C and pepsin C of Japanese monkey were each found to give a single precipitin line which fused completely with each other on agarose plate. On the other hand, cathepsin D-I purified from the monkey lung, and pepsinogens A (I, II, III-1, III-2 and III-3) obtained from the monkey gastric mucosa failed to precipitate with the antiserum. With the antiserum against the monkey pepsinogen C, the same results were obtained. Further, procathepsin D-II and pepsinogen C were shown to have the same amino-terminal amino acid sequence, Ala-Val-Val-Lys-Val-Pro-Leu-Lys-Lys-Phe-Lys-. All these results indicate a strong similarity of procathepsin D-II and cathepsin D-II to pepsinogen C and pepsin C, respectively.
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PMID:Identification of monkey lung procathepsin D-II as a pepsinogen-C-like acid protease zymogen. 640 25

Two types of cathepsin D (cathepsins D-I and D-II) were purified from rhesus monkey lung to homogeneity as judged from disc gel electrophoresis. Cathepsin D-I was purified about 2,000-fold with a 5.1% yield while cathepsin D-II was purified about 2,300-fold with a 14.3% yield. Both cathepsins D were rich in the lysosome fraction of the lung, but appeared to be present in part extracellularly. Both showed a molecular weight of about 35,000 on Sephadex G-100 chromatography, and about 41,000 on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Cathepsin D-I showed the maximal activity on bovine hemoglobin and albumin at pH 3.4 and 4.0, respectively. It was most stable in the pH range of 5 to 7, but was rather unstable outside this pH range. Cathepsin D-II was quite similar in properties to that from Japanese monkey lung (Moriyama, A. & Takahashi, K. (1978) J. Biochem. 83, 441-451), and was remarkably stable in the pH range of 1-9. Under the conditions used, it retained at least 80% of the original activity when incubated at 37 degrees C for 20 h in this pH range. This stability seems to allow cathepsin D-II to be fairly active even at pH 1.0. Both cathepsins D acted on protein substrates fairly similarly and hydrolyzed hemoglobin most rapidly among the proteins tested. They did not hydrolyze N-acetyl-L-phenylalanyl-3,5-diiodotyrosine. Upon incubation with the oxidized B-chain of insulin, both cathepsins D hydrolyzed the Ala-Leu, Leu-Tyr, Tyr-Leu, Phe-Phe, and Phe-Tyr bonds at both pH 3.0 and 5.0. In addition, cathepsin D-II hydrolyzed the Leu-Val and Tyr-Thr bonds at pH 3.0 and the Val-Asn bond at pH 5.0. Both cathepsins D were inactivated by acid protease-specific inhibitors such as pepstatin, 1,2-epoxy-3-(p-nitrophenoxy)propane, p-bromophenacyl bromide, and diazoacetyl-DL-norleucine methyl ester, although cathepsin D-II was much less susceptible to these reagents except p-bromophenacyl bromide.
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PMID:Cathepsins D from rhesus monkey lung. Purification and characterization. 699 76

An acid proteinase of Dirofilaria immitis worms was purified 437-fold by gel filtration on Sephadex G-75 followed by pepstatin-Agarose gel affinity chromatography. The enzyme with a molecular weight of 42 kDa was homogeneous as judged by both affinity chromatography and SDS-polyacrylamide gel electrophoresis. Polyacrylamide disc electrophoresis at pH 8.9, however, revealed that the enzyme was composed of five multi-forms, all carrying proteinase activity. Optimum pH of the enzyme was in the range of pH 2.8 to 3.4, and its isoelectric point ranged between 5.8 and 6.4. The purified proteinase showed a potent activity against hemoglobin and myoglobin releasing acid soluble peptides, but not free amino acids. When enzymatic properties of the proteinase was compared with mammalian cathepsin D and pepsin, D. immitis proteinase activity was reduced to about 80% of the initial activity by incubating at neutral pH and 50 degrees C for 5 min, just like cathepsin D, which remained intact. Pepsin activity was completely destroyed under the same condition. An aspartic proteinase inhibitor, 1,2-epoxy-3-(p-nitrophenoxy)propane, which inhibited pepsin by 30% at 37 degrees C for 10 min, did show little effects on D. immitis proteinase and cathepsin D. Inhibitory effect of diazoacetyl-DL-norleucine methyl ester (DAN) on D. immitis proteinase was intermediate (50% after 60 min). Immunolocalization of the proteinase in the worm tissue using its monoclonal antibodies revealed that the enzyme was localized in the intestine as well as uterine wall and some small granules of microfilariae in the uterus.
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PMID:Purification and characterization of an acid proteinase from Dirofilaria immitis worms. 854 Mar 32

The host specificity of Ralstonia solanacearum, the causal organism of bacterial wilt on many solanaceous crops, is poorly understood. To identify a gene conferring host specificity of the bacterium, SL341 (virulent to hot pepper but avirulent to potato) and SL2029 (virulent to potato but avirulent to hot pepper) were chosen as representative strains. We identified a gene, rsa1, from SL2029 that confers avirulence to SL341 in hot pepper. The rsa1 gene encoding an 11.8-kDa protein possessed the perfect consensus hrp(II) box motif upstream of the gene. Although the expression of rsa1 was activated by HrpB, a transcriptional activator for hrp gene expression, Rsa1 protein was secreted in an Hrp type III secretion-independent manner. Rsa1 exhibited weak homology with an aspartic protease, cathepsin D, and possessed protease activity. Two specific aspartic protease inhibitors, pepstatin A and diazoacetyl-d,l-norleucine methyl ester, inhibited the protease activity of Rsa1. Substitution of two aspartic acid residues with alanine at positions 54 and 59 abolished protease activity. The SL2029 rsa1 mutant was much less virulent than the wild-type strain, but did not induce disease symptoms in hot pepper. These data indicate that Rsa1 is an extracellular aspartic protease and plays an important role for the virulence of SL2029 in potato.
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PMID:An HrpB-dependent but type III-independent extracellular aspartic protease is a virulence factor of Ralstonia solanacearum. 2145 32