Further characteristics of cathepsin D inhibitor from potatoes

H. Keilová; V. Tomášek

doi:10.1135/cccc19762440

Review for "Expression profiling of precuneus layer III cathepsin D‐immunopositive pyramidal neurons in mild cognitive impairment and Alzheimer's disease: Evidence for neuronal signaling vulnerability"

10.1002/cne.24929/v2/review1 ◽

2020 ◽

Keyword(s):

Alzheimer’S Disease ◽

Alzheimer's Disease ◽

Cognitive Impairment ◽

Mild Cognitive Impairment ◽

Expression Profiling ◽

Cathepsin D ◽

Pyramidal Neurons ◽

Neuronal Signaling

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644: Cathepsin D is a Marker for Early Stage Renal Cell Cancer and Predicts the Presence of Distant Metastases

The Journal of Urology ◽

10.1016/s0022-5347(18)34884-5 ◽

2005 ◽

Vol 173 (4S) ◽

pp. 175-175

Author(s):

Axel S. Merseburger ◽

Joerg Hennenlotter ◽

Perikles Simon ◽

Marcus Horstmann ◽

Arnulf Stenzl ◽

...

Keyword(s):

Renal Cell Cancer ◽

Renal Cell ◽

Cathepsin D ◽

Early Stage ◽

Cell Cancer ◽

Distant Metastases

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Increased Cathepsin D expression has a negative prognostic effect on survival in gemcitabine treated patients with pancreatic ductal adenocarcinoma

Zeitschrift für Gastroenterologie ◽

10.1055/s-0038-1668984 ◽

2018 ◽

Vol 56 (08) ◽

pp. e324-e325

Author(s):

UM Mahajan ◽

E Goni ◽

E Langhoff ◽

S Kruger ◽

S Ormanns ◽

...

Keyword(s):

Pancreatic Ductal Adenocarcinoma ◽

Cathepsin D ◽

Ductal Adenocarcinoma ◽

Prognostic Effect

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Faculty Opinions recommendation of Requirement of the human GARP complex for mannose 6-phosphate-receptor-dependent sorting of cathepsin D to lysosomes.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.718529687.793498665 ◽

2014 ◽

Author(s):

Paul Gleeson

Keyword(s):

Cathepsin D ◽

Mannose 6 Phosphate ◽

Garp Complex

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Effect of Temperature and pH on the Secondary Structure and Denaturation Process of Jumbo Squid Hepatopancreas Cathepsin D.

Protein and Peptide Letters ◽

10.2174/0929866526666190405124353 ◽

2019 ◽

Vol 26 (7) ◽

pp. 532-541 ◽

Cited By ~ 1

Author(s):

Cadena-Cadena Francisco ◽

Cárdenas-López José Luis ◽

Ezquerra-Brauer Josafat Marina ◽

Cinco-Moroyoqui Francisco Javier ◽

López-Zavala Alonso Alexis ◽

...

Keyword(s):

Circular Dichroism ◽

Secondary Structure ◽

Cathepsin D ◽

Thermal Denaturation ◽

Protein Denaturation ◽

Marine Organisms ◽

Effect Of Temperature ◽

Jumbo Squid ◽

Α Helix ◽

Circular Dichroïsm

Background: Cathepsin D is a lysosomal enzyme that is found in all organisms acting in protein turnover, in humans it is present in some types of carcinomas, and it has a high activity in Parkinson's disease and a low activity in Alzheimer disease. In marine organisms, most of the research has been limited to corroborate the presence of this enzyme. It is known that cathepsin D of some marine organisms has a low thermostability and that it has the ability to have activity at very acidic pH. Cathepsin D of the Jumbo squid (Dosidicus gigas) hepatopancreas was purified and partially characterized. The secondary structure of these enzymes is highly conserved so the role of temperature and pH in the secondary structure and in protein denaturation is of great importance in the study of enzymes. The secondary structure of cathepsin D from jumbo squid hepatopancreas was determined by means of circular dichroism spectroscopy. Objective: In this article, our purpose was to determine the secondary structure of the enzyme and how it is affected by subjecting it to different temperature and pH conditions. Methods: Circular dichroism technique was used to measure the modifications of the secondary structure of cathepsin D when subjected to different treatments. The methodology consisted in dissecting the hepatopancreas of squid and freeze drying it. Then a crude extract was prepared by mixing 1: 1 hepatopancreas with assay buffer, the purification was in two steps; the first step consisted of using an ultrafiltration membrane with a molecular cut of 50 kDa, and the second step, a pepstatin agarose resin was used to purification the enzyme. Once the enzyme was purified, the purity was corroborated with SDS PAGE electrophoresis, isoelectric point and zymogram. Circular dichroism is carried out by placing the sample with a concentration of 0.125 mg / mL in a 3 mL quartz cell. The results were obtained in mdeg (millidegrees) and transformed to mean ellipticity per residue, using 111 g/mol molecular weight/residue as average. Secondary-structure estimation from the far-UV CD spectra was calculated using K2D Dichroweb software. Results: It was found that α helix decreases at temperatures above 50 °C and above pH 4. Heating the enzyme above 70°C maintains a low percentage of α helix and increases β sheet. Far-UV CD measurements of cathepsin D showed irreversible thermal denaturation. The process was strongly dependent on the heating rate, accompanied by a process of oligomerization of the protein that appears when the sample is heated, and maintained a certain time at this temperature. An amount typically between 3 and 4% α helix of their secondary structure remains unchanged. It is consistent with an unfolding process kinetically controlled due to the presence of an irreversible reaction. The secondary structure depends on pH, and a pH above 4 causes α helix structures to be modified. Conclusion: In conclusion, cathepsin D from jumbo squid hepatopancreas showed retaining up to 4% α helix at 80°C. The thermal denaturation of cathepsin D at pH 3.5 is under kinetic control and follows an irreversible model.

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