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Purification, crystallization and preliminary X-ray diffraction experiment of nattokinase from Bacillus subtilis natto

Abstract

Nattokinase, a protein found in high levels in the traditional Japanese food natto, has been reported to have high thrombolytic activity. In the present study, the crystallization of native nattokinase and the collection of X-ray diffraction date from a nattokinase crystal to a resolution of 1.74 Å are reported. Nattokinase is a single polypeptide chain composed of 275 amino acids (molecular weight 27 724) which displays strong fibrinolytic activity. Moreover, it can activate other fibrinolytic enzymes such as pro-urokinase and tissue plasminogen activator. In the present study, native nattokinase from Bacillus subtilis natto was purified using gel-filtration chromatography and crystallized to give needle-like crystals which could be used for X-ray diffraction experiments. The crystals belonged to space group C2, with unit-cell parameters a = 74.3, b = 49.9, c = 56.3 Å, β = 95.2°. Diffraction images were processed to a resolution of 1.74 Å with an R{sub merge} of 5.2% (15.3% in the highest resolution shell) and a completeness of 69.8% (30.0% in the highest resolution shell). This study reports the first X-ray diffraction analysis of nattokinase.
Authors:
Yanagisawa, Yasuhide; [1]  Chatake, Toshiyuki; [2]  Chiba-Kamoshida, Kaori; [3]  Naito, Sawa; Ohsugi, Tadanori; Sumi, Hiroyuki; [4]  Yasuda, Ichiro; [1]  Morimoto, Yukio; [2]  Faculty of Pharmaceutical Sciences, Chiba Institute of Science, 15-8 Shiomi-cho, Choshi, Chiba 288-0025 (Japan)]
  1. Faculty of Pharmaceutical Sciences, Chiba Institute of Science, 15-8 Shiomi-cho, Choshi, Chiba 288-0025 (Japan)
  2. Research Reactor Institute, Kyoto University, Asashironishi 2, Kumatori, Sennan, Osaka 590-0494 (Japan)
  3. National Institute of Advanced Industrial Science and Technology (AIST), Umezono 1-1-1, Tsukuba, Ibaraki 305-8568 (Japan)
  4. Kurashiki University of Science and Arts, Nishinoura 2640, Tsurajima-cho, Kurashiki, Okayama 712-8505 (Japan)
Publication Date:
Dec 01, 2010
Product Type:
Journal Article
Resource Relation:
Journal Name: Acta Crystallographica. Section F; Journal Volume: 66; Journal Issue: Pt 12; Other Information: PMCID: PMC2998380; PMID: 21139221; PUBLISHER-ID: fw5279; OAI: oai:pubmedcentral.nih.gov:2998380; Copyright (c) International Union of Crystallography 2010; Country of input: International Atomic Energy Agency (IAEA)
Subject:
75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY; CRYSTALLIZATION; CRYSTALS; GELS; IMAGES; MOLECULAR WEIGHT; RESOLUTION; SHELLS; SPACE GROUPS; X-RAY DIFFRACTION
OSTI ID:
22367377
Country of Origin:
United Kingdom
Language:
English
Other Identifying Numbers:
Journal ID: ISSN 1744-3091; CODEN: ACSFCL; Other: PII: S1744309110043137; TRN: GB15$1130072709
Availability:
Available from http://dx.doi.org/10.1107/S1744309110043137; Available from http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2998380
Submitting Site:
INIS
Size:
page(s) 1670-1673
Announcement Date:
Jul 29, 2015

Citation Formats

Yanagisawa, Yasuhide, Chatake, Toshiyuki, Chiba-Kamoshida, Kaori, Naito, Sawa, Ohsugi, Tadanori, Sumi, Hiroyuki, Yasuda, Ichiro, Morimoto, Yukio, and Faculty of Pharmaceutical Sciences, Chiba Institute of Science, 15-8 Shiomi-cho, Choshi, Chiba 288-0025 (Japan)]. Purification, crystallization and preliminary X-ray diffraction experiment of nattokinase from Bacillus subtilis natto. United Kingdom: N. p., 2010. Web. doi:10.1107/S1744309110043137.
Yanagisawa, Yasuhide, Chatake, Toshiyuki, Chiba-Kamoshida, Kaori, Naito, Sawa, Ohsugi, Tadanori, Sumi, Hiroyuki, Yasuda, Ichiro, Morimoto, Yukio, & Faculty of Pharmaceutical Sciences, Chiba Institute of Science, 15-8 Shiomi-cho, Choshi, Chiba 288-0025 (Japan)]. Purification, crystallization and preliminary X-ray diffraction experiment of nattokinase from Bacillus subtilis natto. United Kingdom. https://doi.org/10.1107/S1744309110043137
Yanagisawa, Yasuhide, Chatake, Toshiyuki, Chiba-Kamoshida, Kaori, Naito, Sawa, Ohsugi, Tadanori, Sumi, Hiroyuki, Yasuda, Ichiro, Morimoto, Yukio, and Faculty of Pharmaceutical Sciences, Chiba Institute of Science, 15-8 Shiomi-cho, Choshi, Chiba 288-0025 (Japan)]. 2010. "Purification, crystallization and preliminary X-ray diffraction experiment of nattokinase from Bacillus subtilis natto." United Kingdom. https://doi.org/10.1107/S1744309110043137.
@misc{etde_22367377,
title = {Purification, crystallization and preliminary X-ray diffraction experiment of nattokinase from Bacillus subtilis natto}
author = {Yanagisawa, Yasuhide, Chatake, Toshiyuki, Chiba-Kamoshida, Kaori, Naito, Sawa, Ohsugi, Tadanori, Sumi, Hiroyuki, Yasuda, Ichiro, Morimoto, Yukio, and Faculty of Pharmaceutical Sciences, Chiba Institute of Science, 15-8 Shiomi-cho, Choshi, Chiba 288-0025 (Japan)]}
abstractNote = {Nattokinase, a protein found in high levels in the traditional Japanese food natto, has been reported to have high thrombolytic activity. In the present study, the crystallization of native nattokinase and the collection of X-ray diffraction date from a nattokinase crystal to a resolution of 1.74 Å are reported. Nattokinase is a single polypeptide chain composed of 275 amino acids (molecular weight 27 724) which displays strong fibrinolytic activity. Moreover, it can activate other fibrinolytic enzymes such as pro-urokinase and tissue plasminogen activator. In the present study, native nattokinase from Bacillus subtilis natto was purified using gel-filtration chromatography and crystallized to give needle-like crystals which could be used for X-ray diffraction experiments. The crystals belonged to space group C2, with unit-cell parameters a = 74.3, b = 49.9, c = 56.3 Å, β = 95.2°. Diffraction images were processed to a resolution of 1.74 Å with an R{sub merge} of 5.2% (15.3% in the highest resolution shell) and a completeness of 69.8% (30.0% in the highest resolution shell). This study reports the first X-ray diffraction analysis of nattokinase.}
doi = {10.1107/S1744309110043137}
journal = []
issue = {Pt 12}
volume = {66}
journal type = {AC}
place = {United Kingdom}
year = {2010}
month = {Dec}
}