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Title: X-ray driven reduction of Cpd I of Catalase-3 from N. crassa reveals differential sensitivity of active sites and formation of ferrous state

Abstract

Catalases are biotechnologically relevant enzymes due to their applications in food technology, bioremediation, and biomedicine. The dismutation of hydrogen peroxide occurs in two steps; in the first one, the enzyme forms an oxidized compound I (Cpd I) and in the second one, the enzyme is reduced to the ferric state. In this research work, we analyzed the reduction of Cpd I by X-ray radiation damage during diffraction experiments in crystals of CAT-3, a Large-Size Subunit Catalase (LSC) from Neurospora crassa. A Multi-Crystal Data collection Strategy was applied in order to obtain the Cpd I structure at a resolution of 2.2 Å; this intermediate was highly sensitive to X-ray and was easily reduced at very low deposited radiation dose, causing breakage of the Fe=O bond. The comparison of the structures reflected reduced intermediates and also evidenced the differential sensitivity per monomer. The resting ferric state was reduced to the ferrous state, an intermediate without a previous report in LSC. The chemically obtained Cpd I and the X-ray reduced intermediates were identified by UV-visible microspectrometry coupled to data collection. The differential sensitivity and the formation of a ferrous state are discussed, emphasizing the importance of the correct interpretation in the oxidation statemore » of the iron heme.« less

Authors:
 [1];  [2];  [3];  [4];  [5]
  1. Universidad Nacional Autónoma de México, Ensenada, B. C. (Mexico)
  2. Brookhaven National Lab. (BNL), Upton, NY (United States)
  3. Stanford Univ., Menlo Park, CA (United States). Stanford Synchrotron Radiation LIghtsource.
  4. Universidad Nacional Autónoma de México, Ciudad de México (Mexico)
  5. Universidad Nacional Autónoma de México, Cuernavaca, Morelos (Mexico)
Publication Date:
Research Org.:
SLAC National Accelerator Lab., Menlo Park, CA (United States); Brookhaven National Lab. (BNL), Upton, NY (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22); USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23); National Institutes of Health (NIH); National Institute of General Medical Sciences
OSTI Identifier:
1532492
Alternate Identifier(s):
OSTI ID: 1546050; OSTI ID: 1547569
Report Number(s):
BNL-211925-2019-JAAM
Journal ID: ISSN 0003-9861
Grant/Contract Number:  
PAPIIT IN204611; GM-0080; AC02-98CH10886; AC02-76SF00515; SC0012704
Resource Type:
Accepted Manuscript
Journal Name:
Archives of Biochemistry and Biophysics
Additional Journal Information:
Journal Volume: 666; Journal Issue: C; Journal ID: ISSN 0003-9861
Publisher:
Elsevier
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; Catalase; Composite datasets; Compound I; Ferrous heme; X-ray reduction; 36 MATERIALS SCIENCE

Citation Formats

Zárate-Romero, Andrés, Stojanoff, Vivian, Cohen, Aina E., Hansberg, Wilhelm, and Rudiño-Piñera, Enrique. X-ray driven reduction of Cpd I of Catalase-3 from N. crassa reveals differential sensitivity of active sites and formation of ferrous state. United States: N. p., 2019. Web. doi:10.1016/j.abb.2019.03.020.
Zárate-Romero, Andrés, Stojanoff, Vivian, Cohen, Aina E., Hansberg, Wilhelm, & Rudiño-Piñera, Enrique. X-ray driven reduction of Cpd I of Catalase-3 from N. crassa reveals differential sensitivity of active sites and formation of ferrous state. United States. doi:10.1016/j.abb.2019.03.020.
Zárate-Romero, Andrés, Stojanoff, Vivian, Cohen, Aina E., Hansberg, Wilhelm, and Rudiño-Piñera, Enrique. Sat . "X-ray driven reduction of Cpd I of Catalase-3 from N. crassa reveals differential sensitivity of active sites and formation of ferrous state". United States. doi:10.1016/j.abb.2019.03.020.
@article{osti_1532492,
title = {X-ray driven reduction of Cpd I of Catalase-3 from N. crassa reveals differential sensitivity of active sites and formation of ferrous state},
author = {Zárate-Romero, Andrés and Stojanoff, Vivian and Cohen, Aina E. and Hansberg, Wilhelm and Rudiño-Piñera, Enrique},
abstractNote = {Catalases are biotechnologically relevant enzymes due to their applications in food technology, bioremediation, and biomedicine. The dismutation of hydrogen peroxide occurs in two steps; in the first one, the enzyme forms an oxidized compound I (Cpd I) and in the second one, the enzyme is reduced to the ferric state. In this research work, we analyzed the reduction of Cpd I by X-ray radiation damage during diffraction experiments in crystals of CAT-3, a Large-Size Subunit Catalase (LSC) from Neurospora crassa. A Multi-Crystal Data collection Strategy was applied in order to obtain the Cpd I structure at a resolution of 2.2 Å; this intermediate was highly sensitive to X-ray and was easily reduced at very low deposited radiation dose, causing breakage of the Fe=O bond. The comparison of the structures reflected reduced intermediates and also evidenced the differential sensitivity per monomer. The resting ferric state was reduced to the ferrous state, an intermediate without a previous report in LSC. The chemically obtained Cpd I and the X-ray reduced intermediates were identified by UV-visible microspectrometry coupled to data collection. The differential sensitivity and the formation of a ferrous state are discussed, emphasizing the importance of the correct interpretation in the oxidation state of the iron heme.},
doi = {10.1016/j.abb.2019.03.020},
journal = {Archives of Biochemistry and Biophysics},
number = C,
volume = 666,
place = {United States},
year = {2019},
month = {3}
}

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This content will become publicly available on March 30, 2020
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