6O44

Insight into subtilisin E-S7 cleavage pattern based on crystal structure and hydrolysates peptide analysis

Summary for 6O44

• Entry DOI: 10.2210/pdb6o44/pdb
• Descriptor: Nattokinase, CALCIUM ION, SULFATE ION, ... (5 entities in total)
• Functional Keywords: subtilisin, hydrolase
• Biological source: Bacillus subtilis
• Total number of polymer chains: 2
• Total formula weight: 58354.37

Authors

Tang, H.,Shi, K.,Aihara, H. (deposition date: 2019-02-28, release date: 2019-04-10, Last modification date: 2023-10-11)

Primary citation

Tang, H.,Zhang, J.,Shi, K.,Aihara, H.,Du, G.
Insight into subtilisin E-S7 cleavage pattern based on crystal structure and hydrolysates peptide analysis.
Biochem. Biophys. Res. Commun., 512:623-628, 2019

PubMed Abstract: The X-ray crystallographic structure of the mature form of subtilisin E-S7 (SES7) at 1.90 Å resolution is reported here. Structural comparisons between the previously reported propeptide-subtilisin E complex (1SCJ) and our mature form subtilisin E-S7 (6O44) provide insight into active site adjustments involved in catalysis and specificity. To further investigate the protease substrate selectivity mechanism, we used SES7 to hydrolyze skim milk and analyzed the hydrolysates by LC-MS for peptide identification. The cleavage pattern suggests a high preference for proline at substrate P2 position. The results based on the peptide analysis are consistent with our structural observations, which is instrumental in future protein engineering by rational design. Furthermore, the ACE-inhibitor and NLN-inhibitor activity of the hydrolysates were determined to assess the utility of SES7 for further industrial applications; IC-ACE = 67 ± 0.92 μg/mL and IC-NLN = 263 ± 13 μg/mL.

PubMed: 30914195
DOI: 10.1016/j.bbrc.2019.03.064

Experimental method

X-RAY DIFFRACTION (1.83 Å)