Studies on Pituitary Catkepsin D with Artificial
Substrates

Reinharz, A.; Roth, Marc

doi:10.1159/000459571

Cited by 20 publications

(3 citation statements)

References 11 publications

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“…10 presents the reaction of amino acids with fluorescamine. The radioelements used for hemoglobin labeling include 3 H, 14 C, 125 I and 131 I [90][91][92]. Addition of a pigment or a fluorizing compound causes substantial changes in globin structure and is frequently associated with reduced susceptibility to cathepsin D action.…”

Section: Native Denaturated and Labeled Proteinsmentioning

confidence: 99%

Quantitative determination and localization of cathepsin D and its inhibitors.

Minarowska

Karwowska²,

Gacko³

2009

Folia Histochem Cytobiol.

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Cathepsin D (EC 3.4.23.5) is a lysosomal aspartyl endopeptidase, localized in all cells and tissues, except for mature erythrocytes [1][2][3]. Methods used to determine the activity, concentration and cellular distribution of cathepsin D, but not its inhibitors, have previously been the subject of literature reports [4][5][6][7][8][9][10][11][12]. However, since the time of their publication a number of new substrates and analytical techniques have been implemented. Structure, specificity, mechanism of actionCathepsin D is synthesized in the rough endoplasmic reticulum as preprocathepsin D, built up of 412 amino acid residues [13][14][15]. As a result of cleavage of the 20-amino acid signal prepeptide, it is converted into procathepsin D which undergoes glycosylation and disulphide bridges are formed in its molecule. Procathepsin D is transported from cisterns of the rough endoplasmic reticulum to the Golgi apparatus, from which, with the involvement of mannoso-6-phosphate (M-P-6) receptors, it is transferred to primary lysosomes [16,17]. As the M-6-P receptors are known to occur in the primary lysosomes but not in the mature ones, they can be used to distinguish between these two types of lysosomes [18]. In the acidic environment of the lysosomes (pH 4.5-5.5), due to autocatalytic cleavage of the 44-amino acid propeptide from the N-terminal molecule, procathepsin D is converted into the active one-chain form. The actions of cysteine proteinase, aminopeptidases and carboxypeptidases lead to the formation of an active two-chain form of cathepsin D (Fig. 1). These chains are bound by hydrophobic bonds. The molecular weight of the ultimate mature form of cathepsin D is 48 (14+34) kDa. The proteolytic activities of the one-chain and two-chain forms are very similar [19,20]. Modification of the polypeptide chain, different oligosaccharide composition types and phosphorylation/dephosphorylation in the amino saccharide residues contribute to marked molecular heterogenicity of cathepsin D and cause differences in isoelectric points of the respective isoenzymes between pH 4.5 -6.5 [21,22].The use of peptides with the known primary structure allows identification of amino acid residues that form peptide bonds cleaved by cathepsin D. For this purpose, synthetic peptides [23] and chains A and B of bovine insulin can be used (Fig. 2). Cathepsin D cleaves the peptide bonds found within the polypeptide chain, formed by carboxyl groups of the hydrophobic amino acid residues: aromatic -trypto-

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Section: Native Denaturated and Labeled Proteinsmentioning

confidence: 99%

Quantitative determination and localization of cathepsin D and its inhibitors.

Minarowska

Karwowska²,

Gacko³

2009

Folia Histochem Cytobiol.

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“…hemoglobin (Hb) and BSA. Cathepsin D incubation with Z8PNA was performed according to (he method of Reinharz and Roth [25], using a 50 m M citrate-phosphate buffer at pH 3.8.…”

Section: Assay O F Cathepsin D Activitymentioning

confidence: 99%

Renin-Like and Cathepsin D Activities in Bovine Pineal Glands

Agostini¹,

Reinharz²,

Vallotton³

1982

Neuroendocrinology

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Pineal extract is shown to contain both renin-like and cathepsin D activities. Evidence of renin-like activity in the bovine pineal gland was brought by incubation with natural and synthetic renin substrates and by inhibition by pepstatin. Cathepsin D activity was demonstrated by incubation with hemoglobin and synthetic ñuorogenic peptide. The separation of both activities was performed by affinity chromatography on a caseinyl-Sepharose gel. The elution of the extract on affinity chromatography allowed to separate the renin-like activity, which is not retained by the gel at acid pH, from cathepsin D activity, which binds to the column at acid pH and is eluted at alkaline pH. The isolated pineal renin-like activity was found higher on tetradecapeptide renin substrate than on angiotensinogen at pH 5.5. The pH dependence of pineal renin-like activity showed two peaks of activity. One broad peak between pH 6 and 8 and one sharp peak at pH 3.5–4. These results demonstrate the existence of renin-like and cathepsin D activities in bovine pineal gland. They suggest moreover that the renin-like activity measured might represent a mixture of at least two different enzymatic activities.

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“…There has been considerable interest surrounding the possibility that extrarenal sources of renin exist and perhaps are important in physiological regulation (Skeggs, Lentz, Kahn, Doer & Levine, 1969;Reinharz & Roth, 1971; Day & Reid, 1976;Hackenthal, Hackenthal & Hilgenfeldt, 1978). The presence of renin has been inferred by demonstrating that tissue homogenates can 14.…”

Section: Vascular Wall Reninmentioning

confidence: 99%

Specific Inhibitors of Renin

Haber

1980

Clinical Science

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Studies on Pituitary Catkepsin D with Artificial Substrates

Cited by 20 publications

References 11 publications

Quantitative determination and localization of cathepsin D and its inhibitors.

Quantitative determination and localization of cathepsin D and its inhibitors.

Renin-Like and Cathepsin D Activities in Bovine Pineal Glands

Specific Inhibitors of Renin

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