Cargando…

Crystallographic analysis and phasing of iron-assimilating protein 1 (FEA1) from Chlamydomonas reinhardtii

As an essential component of protein cofactors, iron is important for all photosynthetic organisms. In Chlamydomonas reinhardtii, iron levels are strictly controlled by proteins such as iron-assimilating protein 1 (FEA1). This periplasmic protein is expressed under conditions of iron deficiency, but...

Descripción completa

Detalles Bibliográficos
Autores principales: Juniar, Linda, Adlfar, Vida, Hippler, Michael, Tanaka, Hideaki, Kurisu, Genji
Formato: Online Artículo Texto
Lenguaje:English
Publicado: International Union of Crystallography 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8098128/
https://www.ncbi.nlm.nih.gov/pubmed/33949973
http://dx.doi.org/10.1107/S2053230X21003952
_version_ 1783688441869893632
author Juniar, Linda
Adlfar, Vida
Hippler, Michael
Tanaka, Hideaki
Kurisu, Genji
author_facet Juniar, Linda
Adlfar, Vida
Hippler, Michael
Tanaka, Hideaki
Kurisu, Genji
author_sort Juniar, Linda
collection PubMed
description As an essential component of protein cofactors, iron is important for all photosynthetic organisms. In Chlamydomonas reinhardtii, iron levels are strictly controlled by proteins such as iron-assimilating protein 1 (FEA1). This periplasmic protein is expressed under conditions of iron deficiency, but its mechanisms of function remain unknown. Because FEA1 has no amino-acid similarity to protein structures in the Protein Data Bank, its crystal structure cannot be solved by molecular replacement. Here, recombinant FEA1 protein lacking the N-terminal signal sequence was successfully purified and crystals of apo FEA1 were obtained by hanging-drop vapor diffusion. Neither selenomethionine substitution nor heavy-atom derivatization was successful; therefore, the phase problem of FEA1 crystals was solved by the native sulfur SAD method using long-wavelength X-rays (2.7 Å). Laser-cutting technology was used to increase the signal-to-noise ratio and derive an initial structure. This study will lead to further structural studies of FEA1 to understand its function and its links to the iron-assimilation pathway.
format Online
Article
Text
id pubmed-8098128
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher International Union of Crystallography
record_format MEDLINE/PubMed
spelling pubmed-80981282021-05-18 Crystallographic analysis and phasing of iron-assimilating protein 1 (FEA1) from Chlamydomonas reinhardtii Juniar, Linda Adlfar, Vida Hippler, Michael Tanaka, Hideaki Kurisu, Genji Acta Crystallogr F Struct Biol Commun Research Communications As an essential component of protein cofactors, iron is important for all photosynthetic organisms. In Chlamydomonas reinhardtii, iron levels are strictly controlled by proteins such as iron-assimilating protein 1 (FEA1). This periplasmic protein is expressed under conditions of iron deficiency, but its mechanisms of function remain unknown. Because FEA1 has no amino-acid similarity to protein structures in the Protein Data Bank, its crystal structure cannot be solved by molecular replacement. Here, recombinant FEA1 protein lacking the N-terminal signal sequence was successfully purified and crystals of apo FEA1 were obtained by hanging-drop vapor diffusion. Neither selenomethionine substitution nor heavy-atom derivatization was successful; therefore, the phase problem of FEA1 crystals was solved by the native sulfur SAD method using long-wavelength X-rays (2.7 Å). Laser-cutting technology was used to increase the signal-to-noise ratio and derive an initial structure. This study will lead to further structural studies of FEA1 to understand its function and its links to the iron-assimilation pathway. International Union of Crystallography 2021-04-28 /pmc/articles/PMC8098128/ /pubmed/33949973 http://dx.doi.org/10.1107/S2053230X21003952 Text en © Juniar et al. 2021 https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution (CC-BY) Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are cited.
spellingShingle Research Communications
Juniar, Linda
Adlfar, Vida
Hippler, Michael
Tanaka, Hideaki
Kurisu, Genji
Crystallographic analysis and phasing of iron-assimilating protein 1 (FEA1) from Chlamydomonas reinhardtii
title Crystallographic analysis and phasing of iron-assimilating protein 1 (FEA1) from Chlamydomonas reinhardtii
title_full Crystallographic analysis and phasing of iron-assimilating protein 1 (FEA1) from Chlamydomonas reinhardtii
title_fullStr Crystallographic analysis and phasing of iron-assimilating protein 1 (FEA1) from Chlamydomonas reinhardtii
title_full_unstemmed Crystallographic analysis and phasing of iron-assimilating protein 1 (FEA1) from Chlamydomonas reinhardtii
title_short Crystallographic analysis and phasing of iron-assimilating protein 1 (FEA1) from Chlamydomonas reinhardtii
title_sort crystallographic analysis and phasing of iron-assimilating protein 1 (fea1) from chlamydomonas reinhardtii
topic Research Communications
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8098128/
https://www.ncbi.nlm.nih.gov/pubmed/33949973
http://dx.doi.org/10.1107/S2053230X21003952
work_keys_str_mv AT juniarlinda crystallographicanalysisandphasingofironassimilatingprotein1fea1fromchlamydomonasreinhardtii
AT adlfarvida crystallographicanalysisandphasingofironassimilatingprotein1fea1fromchlamydomonasreinhardtii
AT hipplermichael crystallographicanalysisandphasingofironassimilatingprotein1fea1fromchlamydomonasreinhardtii
AT tanakahideaki crystallographicanalysisandphasingofironassimilatingprotein1fea1fromchlamydomonasreinhardtii
AT kurisugenji crystallographicanalysisandphasingofironassimilatingprotein1fea1fromchlamydomonasreinhardtii