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Crystallographic analysis of Eisenia hydrolysis-enhancing protein using a long wavelength for native-SAD phasing

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Title: Crystallographic analysis of Eisenia hydrolysis-enhancing protein using a long wavelength for native-SAD phasing
Authors: Sun, Xiaomei Browse this author
Ye, Yuxin Browse this author
Sakurai, Naofumi Browse this author
Kato, Koji Browse this author
Yuasa, Keizo Browse this author
Tsuji, Akihiko Browse this author
Yao, Min Browse this author
Keywords: EHEP
phlorotannin binding
solutionless crystal mount
Issue Date: Jan-2020
Publisher: International Union of Crystallography
Journal Title: Acta crystallographica Section F
Volume: 76
Start Page: 20
End Page: 24
Publisher DOI: 10.1107/S2053230X19016716
Abstract: Eisenia hydrolysis-enhancing protein (EHEP), which is a novel protein that has been identified in Aplysia kurodai, protects beta-glucosidases from phlorotannin inhibition to facilitate the production of glucose from the laminarin abundant in brown algae. Hence, EHEP has attracted attention for its potential applications in producing biofuel from brown algae. In this study, EHEP was purified from the natural digestive fluid of A. kurodai and was crystallized using the sitting-drop vapor-diffusion method. Native and SAD (single-wavelength anomalous diffraction) data sets were successfully collected at resolutions of 1.20 and 2.48 angstrom using wavelengths of 1.0 and 2.1 angstrom, respectively, from crystals obtained in initial screening. The crystals belonged to space group P2(1)2(1)2(1) and contained one EHEP molecule in the asymmetric unit. All 20 S-atom sites in EHEP were located and the phases were determined by the SAD method using the S atoms in the natural protein as anomalous scatterers (native-SAD). After phase improvement, interpretable electron densities were obtained and 58% of the model was automatically built.
Rights: © International Union of Crystallography
Type: article
Appears in Collections:生命科学院・先端生命科学研究院 (Graduate School of Life Science / Faculty of Advanced Life Science) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: 姚 閔

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