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Structural basis of the cystein protease inhibitor Clonorchis sinensis Stefin-1

Journal Article · · Biochemical and Biophysical Research Communications
;  [1];  [2];  [3]
  1. Department of Molecular Biology, College of Natural Sciences, Pusan National University, Jangjeon-dong, Geumjeong-gu, Busan, 46241 (Korea, Republic of)
  2. Pohang Accelerator Laboratory, Pohang University of Science and Technology, 80, Jigok-ro 127 Beon-gil, Nam-gu, Pohang-si, Gyeongsangbuk-do, 37673 (Korea, Republic of)
  3. Department of Parasitology and Tropical Medicine, Institute of Health Sciences, Gyeongsang National University College of Medicine, Jinju, 52727 (Korea, Republic of)

Highlights: • Clonorchis sinensis Stefin-1 has known as a host immune modulator. • The first crystal structure of CsStefin-1 was determined by the MAD method. • CsStefin-1 inhibited CsCF-8 cysteine protease activity. • A molecular docking model of CsStefin-1 and CsCF-8 was developed. • The CsStefin-1 was physically bound to CsCF-8 with an apparent K{sub D} value. Cystein protease plays a critical role as a virulence factor in the development and progression of various diseases. Cystatin is a superfamily of cysteine protease inhibitors that participates in various physiological and pathological processes. The cysteine protease inhibitor CsStein-1 isolated from Clonorchis sinensis belongs to the type 1 stefin of cystatins. This inhibitor regulates the activity and processing of CsCF (Cathepsin F of Clonorchis sienesis), which plays an important role in parasite nutrition and host-parasite interaction. CsStefin-1 has also been proposed as a host immune modulator and a participant in the mechanism associated with anti-inflammatory ability. Here, we report the first crystal structure of CsStefin-1 determined by the multi-wavelength anomalous diffraction (MAD) method to 2.3 Å. There are six molecules of CsStefin-1 per asymmetric unit, with a solvent content of 36.5%. The structure of CsStefin-1 is composed of twisted four-stranded antiparallel β-sheets, a central α-helix, and a short α-helix. We also demonstrate that CsStefin-1 binds to CsCF-8 cysteine protease and inhibits its activity. In addition, a molecular docking model of CsStefin-1 and CsCF-8 was developed using homology modeling based on their structures. The structural information regarding CsStefin-1 and molecular insight into its interaction with CsCF-8 are important to understanding their biological function and to design of inhibitors that modulate cysteine protease activity.

OSTI ID:
23137265
Journal Information:
Biochemical and Biophysical Research Communications, Journal Name: Biochemical and Biophysical Research Communications Journal Issue: 1 Vol. 498; ISSN BBRCA9; ISSN 0006-291X
Country of Publication:
United States
Language:
English

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