Structural basis of cucumisin protease activity regulation by its propeptide

Ami Sotokawauchi, Miyuki Kato-Murayama, Kazutaka Murayama, Toshiaki Hosaka, Iori Maeda, Michio Onjo, Noboru Ohsawa, Dai Ichiro Kato, Kazunari Arima, Mikako Shirouzu

Research output: Contribution to journalArticlepeer-review

6 Citations (Scopus)

Abstract

Cucumisin [EC 3.4.21.25], a subtilisin-like serine endopeptidase, was isolated from melon fruit, Cucumis melo L. Mature cucumisin (67 kDa, 621 residues) is produced by removal of the propeptide (10 kDa, 88 residues) from the cucumisin precursor by subsequence processing. It is reported that cucumisin is inhibited by its own propeptide. The crystal structure of mature cucumisin is reported to be composed of three domains: the subtilisin-like catalytic domain, the protease-associated domain and the C-terminal fibronectin-III-like domain. In this study, the crystal structure of the mature cucumisin•propeptide complex was determined by the molecular replacement method and refined at 1.95 Å resolution. In this complex, the propeptide had a domain of the α-β sandwich motif with four-stranded antiparallel β-sheets, two helices and a strand of the C-terminal region. The β-sheets of the propeptide bind to two parallel surface helices of cucumisin through hydrophobic interaction and 27 hydrogen bonds. The C-terminus of the propeptide binds to the cleft of the active site as peptide substrates. The inhibitory assay suggested that the C-terminal seven residues of the propeptide do not inhibit the cucumisin activity. The crystal structure of the cucumisin•propeptide complex revealed the regulation mechanism of cucumisin activity.

Original languageEnglish
Pages (from-to)45-53
Number of pages9
JournalJournal of biochemistry
Volume161
Issue number1
DOIs
Publication statusPublished - 2017 Jan 1

Keywords

  • Plant serine protease
  • Protease inhibition
  • Protein-propeptide complex
  • Regulation mechanism
  • X-ray crystallography

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Fingerprint Dive into the research topics of 'Structural basis of cucumisin protease activity regulation by its propeptide'. Together they form a unique fingerprint.

Cite this