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

Author(s):

Cadena-Cadena Francisco, Cárdenas-López José Luis*, Ezquerra-Brauer Josafat Marina, Cinco-Moroyoqui Francisco Javier, López-Zavala Alonso Alexis, Santacruz-Ortega Hisila del Carmen and Rivero-Espejel Ignacio Alfredo   Pages 1 - 9 ( 9 )

Abstract:


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. 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.

Keywords:

Jumbo squid, cathepsin D denaturation, circular dichroism, oligomerization, Far-UV CD, hepatopancreas.

Affiliation:

Departamento de Investigación y Posgrado de Alimentos, Universidad de Sonora, Hermosillo, Sonora, Departamento de Investigación y Posgrado de Alimentos, Universidad de Sonora, Hermosillo, Sonora, Departamento de Investigación y Posgrado de Alimentos, Universidad de Sonora, Hermosillo, Sonora, Departamento de Investigación y Posgrado de Alimentos, Universidad de Sonora, Hermosillo, Sonora, Departamento de Ciencias Químico Biológicas, Universidad de Sonora, Hermosillo, Sonora, Departamento de Investigación en Polímeros y Materiales, Universidad de Sonora, Hermosillo, Sonora, Centro de Graduados e Investigación en Química, Instituto Tecnológico de Tijuana, Tijuana, Baja California



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