Characterization and crystal structure of prolyl endopeptidase from abalone (Haliotis discus hannai)
Overview of Li WY et al.
| Authors | Li WY Li Y Chen YL Hu JJ Mengist HM Liu GM Jin T Cao MJ |
|---|---|
| Affiliation | College of Food and Biological Engineering Jimei University Xiamen Fujian 361021 China. Electronic address: mjcao@jmu.edu.cn. |
| Journal | Food Chem |
| Year | 2020 |
Abstract
Aimed to study the characteristics of prolyl endopeptidase (PEP, EC 3.4.21.26) and its possible role in the degradation of collagen, we cloned the full-length cDNA sequence of PEP from abalone (Haliotis discus hannai) (Hdh-PEP). Recombinant Hdh-PEP (rHdh-PEP) was expressed in vitro, its enzymatic properties were detected, and its secondary structure was analyzed by Circular Dichroism (CD). We for the first time determined the 1.5 Å crystal structure of rHdh-PEP. The decomposition effect of rHdh-PEP on collagen peptides was analyzed. Our data revealed that the molecular weight of rHdh-PEP is 85 kDa, consisting of a catalytic domain and a β-propeller domain. The optimal pH and temperature of rHdh-PEP were pH 6.0 and 20 °C, respectively. Using small collagen peptides as substrates, HPLC-ESI-MS analysis confirmed that rHdh-PEP specifically cleaved at the carboxyl side of proline residues, suggesting its role in the degradation of collagen peptides during autolysis.