6O44

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

6O44 の概要

• エントリーDOI: 10.2210/pdb6o44/pdb
• 分子名称: Nattokinase, CALCIUM ION, SULFATE ION, ... (5 entities in total)
• 機能のキーワード: subtilisin, hydrolase
• 由来する生物種: Bacillus subtilis
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 58354.37

構造登録者

Tang, H.,Shi, K.,Aihara, H. (登録日: 2019-02-28, 公開日: 2019-04-10, 最終更新日: 2023-10-11)

主引用文献

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

実験手法

X-RAY DIFFRACTION (1.83 Å)