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Title: Solution structure for an Encephalitozoon cuniculi adrenodoxin-like protein in the oxidized state

Abstract

Encephalitozoon cuniculi is a unicellular, obligate intracellular eukaryotic parasite in the Microsporidia family and one of the agents responsible for microsporidosis infections in humans. Like most Microsporidia, the genome of E. cuniculi is markedly reduced and the organism contains mitochondria-like organelles called mitosomes instead of mitochondria. Here we report the solution NMR structure for a protein physically associated with mitosome-like organelles in E. cuniculi, the 128-residue, adrenodoxin-like protein Ec-Adx (UniProt ID Q8SV19) in the [2Fe-2S] ferredoxin superfamily. Oxidized Ec-Adx contains a mixed four-strand β-sheet, β2-β1-β4-β3 (↓↑↑↓), loosely encircled by three α-helices and two 310-helices. This fold is similar to the structure observed in other adrenodoxin and adrenodoxin-like proteins except for the absence of a fifth anti-parallel β-strand next to β3 and the position of α3. Cross peaks are missing or cannot be unambiguously assigned for 20 amide resonances in the 1H- 15N HSQC spectrum of Ec-Adx. These missing residues are clustered primarily in two regions, G48-V61 and L94-L98, containing the four cysteine residues predicted to ligate the paramagnetic [2Fe-2S] cluster. Missing amide resonances in 1H- 15N HSQC spectra are detrimental to NMR-based solution structure calculations because 1H- 1H NOE restraints are absent (glass half-empty) and this may account for themore » absent β-strand (β5) and the position of α3 in oxidized Ec-Adx. On the other hand, the missing amide resonances unambiguously identify the presence, and immediate environment, of the paramagnetic [2Fe-2S] cluster in oxidized Ec-Adx (glass half-full).« less

Authors:
 [1];  [2];  [3];  [1];  [4];  [5];  [2]; ORCiD logo [6]
  1. Univ. of Washington, Seattle, WA (United States). Dept. of Medicine, Division of Allergy and Infectious Disease, Center for Emerging and Remerging Infectious Disease
  2. Univ. of Washington, Seattle, WA (United States). Dept. of Medicine, Division of Allergy and Infectious Disease, Center for Emerging and Remerging Infectious Disease; Seattle Structural Genomics Center for Infectious Diseases (United States)
  3. Seattle Structural Genomics Center for Infectious Diseases (United States); Seattle Children's Research Inst., Seattle, WA (United States). Center for Global Infectious Disease Research
  4. Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
  5. Seattle Children's Research Institute (United States); Univ. of Washington, Seattle, WA (United States). Dept. of Medicine, Division of Allergy and Infectious Disease, Center for Emerging and Remerging Infectious Disease; Seattle Structural Genomics Center for Infectious Diseases (United States)
  6. Seattle Structural Genomics Center for Infectious Diseases (United States); Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Univ. of Washington, Seattle, WA (United States)
Publication Date:
Research Org.:
Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)
OSTI Identifier:
1599908
Report Number(s):
[PNNL-SA-150252]
[Journal ID: ISSN 0961-8368]
Grant/Contract Number:  
[AC05-76RL01830; HHSH27220120025C; HHSN272200700057C; HHSN272201700059C]
Resource Type:
Accepted Manuscript
Journal Name:
Protein Science
Additional Journal Information:
[ Journal Volume: 29; Journal Issue: 3]; Journal ID: ISSN 0961-8368
Publisher:
The Protein Society
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; electron transport; microspordia; NMR spectroscopy; structural genomics; structure‐based drug discovery

Citation Formats

Shaheen, Shareef, Barrett, Kayleigh F., Subramanian, Sandhya, Arnold, Samuel L. M., Laureanti, Joseph A., Myler, Peter J., Van Voorhis, Wesley C., and Buchko, Garry W. Solution structure for an Encephalitozoon cuniculi adrenodoxin-like protein in the oxidized state. United States: N. p., 2020. Web. doi:10.1002/pro.3818.
Shaheen, Shareef, Barrett, Kayleigh F., Subramanian, Sandhya, Arnold, Samuel L. M., Laureanti, Joseph A., Myler, Peter J., Van Voorhis, Wesley C., & Buchko, Garry W. Solution structure for an Encephalitozoon cuniculi adrenodoxin-like protein in the oxidized state. United States. doi:10.1002/pro.3818.
Shaheen, Shareef, Barrett, Kayleigh F., Subramanian, Sandhya, Arnold, Samuel L. M., Laureanti, Joseph A., Myler, Peter J., Van Voorhis, Wesley C., and Buchko, Garry W. Mon . "Solution structure for an Encephalitozoon cuniculi adrenodoxin-like protein in the oxidized state". United States. doi:10.1002/pro.3818.
@article{osti_1599908,
title = {Solution structure for an Encephalitozoon cuniculi adrenodoxin-like protein in the oxidized state},
author = {Shaheen, Shareef and Barrett, Kayleigh F. and Subramanian, Sandhya and Arnold, Samuel L. M. and Laureanti, Joseph A. and Myler, Peter J. and Van Voorhis, Wesley C. and Buchko, Garry W.},
abstractNote = {Encephalitozoon cuniculi is a unicellular, obligate intracellular eukaryotic parasite in the Microsporidia family and one of the agents responsible for microsporidosis infections in humans. Like most Microsporidia, the genome of E. cuniculi is markedly reduced and the organism contains mitochondria-like organelles called mitosomes instead of mitochondria. Here we report the solution NMR structure for a protein physically associated with mitosome-like organelles in E. cuniculi, the 128-residue, adrenodoxin-like protein Ec-Adx (UniProt ID Q8SV19) in the [2Fe-2S] ferredoxin superfamily. Oxidized Ec-Adx contains a mixed four-strand β-sheet, β2-β1-β4-β3 (↓↑↑↓), loosely encircled by three α-helices and two 310-helices. This fold is similar to the structure observed in other adrenodoxin and adrenodoxin-like proteins except for the absence of a fifth anti-parallel β-strand next to β3 and the position of α3. Cross peaks are missing or cannot be unambiguously assigned for 20 amide resonances in the 1H-15N HSQC spectrum of Ec-Adx. These missing residues are clustered primarily in two regions, G48-V61 and L94-L98, containing the four cysteine residues predicted to ligate the paramagnetic [2Fe-2S] cluster. Missing amide resonances in 1H-15N HSQC spectra are detrimental to NMR-based solution structure calculations because 1H-1H NOE restraints are absent (glass half-empty) and this may account for the absent β-strand (β5) and the position of α3 in oxidized Ec-Adx. On the other hand, the missing amide resonances unambiguously identify the presence, and immediate environment, of the paramagnetic [2Fe-2S] cluster in oxidized Ec-Adx (glass half-full).},
doi = {10.1002/pro.3818},
journal = {Protein Science},
number = [3],
volume = [29],
place = {United States},
year = {2020},
month = {1}
}

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