Development of immunoassays for the iron-regulated proteins ferredoxin and flavodoxin in polar microalgae
While the growth of Southern Ocean phytoplankton is often limited by iron availability, there are no comparable experiments on sea-ice algae. Here we assess the use of ferredoxin and flavodoxin to investigate the iron nutritional status of sea-ice algae and describe the development of a quantitative...
Published in: | Journal of Phycology |
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Language: | English |
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2009
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Online Access: | https://eprints.utas.edu.au/12240/ https://eprints.utas.edu.au/12240/1/DEVELOPMENT_OF_IMMUNOASSAYS_FOR_THE_IRON-REGULATED_PROTEINS_FERREDOXIN_AND_FLAVODOXIN_IN_POLAR_MICROALGAE1.pdf https://doi.org/10.1111/j.1529-8817.2009.00687.x |
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ftunivtasmania:oai:eprints.utas.edu.au:12240 2023-05-15T13:36:47+02:00 Development of immunoassays for the iron-regulated proteins ferredoxin and flavodoxin in polar microalgae Pankowski, A McMinn, A 2009 application/pdf https://eprints.utas.edu.au/12240/ https://eprints.utas.edu.au/12240/1/DEVELOPMENT_OF_IMMUNOASSAYS_FOR_THE_IRON-REGULATED_PROTEINS_FERREDOXIN_AND_FLAVODOXIN_IN_POLAR_MICROALGAE1.pdf https://doi.org/10.1111/j.1529-8817.2009.00687.x en eng https://eprints.utas.edu.au/12240/1/DEVELOPMENT_OF_IMMUNOASSAYS_FOR_THE_IRON-REGULATED_PROTEINS_FERREDOXIN_AND_FLAVODOXIN_IN_POLAR_MICROALGAE1.pdf Pankowski, A and McMinn, A 2009 , 'Development of immunoassays for the iron-regulated proteins ferredoxin and flavodoxin in polar microalgae' , Journal of Phycology, vol. 45 , pp. 771-783 , doi:10.1111/j.1529-8817.2009.00687.x <http://dx.doi.org/10.1111/j.1529-8817.2009.00687.x>. cc_utas ferredoxin flavodoxin immunoassay; iron; microalgae; phytoplankton Article PeerReviewed 2009 ftunivtasmania https://doi.org/10.1111/j.1529-8817.2009.00687.x 2020-05-30T07:26:00Z While the growth of Southern Ocean phytoplankton is often limited by iron availability, there are no comparable experiments on sea-ice algae. Here we assess the use of ferredoxin and flavodoxin to investigate the iron nutritional status of sea-ice algae and describe the development of a quantitative immunoassay for both proteins in marine diatoms. Highaffinity monoclonal antibodies toward both proteins were produced from Cylindrotheca closterium (Ehrenb.) J. M. Lewin et Reimann, and these were used to develop Western blots. Western blots run on whole protein extracts detected both proteins with little cross-reactivity toward other proteins. The two proteins could be successfully quantitated when applied to gels at between 5 and 50 ng in a volume of 25 lL (0.2–2 lg ÆmL)1). Flavodoxin and ferrodoxin expression was examined in the Antarctic diatoms Entomoneis kjellmannii (Cleve) Poulin et Cardinal, Navicula directa (W. Sm.) Ralfs, Fragilariopsis curta (Van Heurck) Hust., Pseudo-nitzschia sp., Porosira glacialis (Grunow) E. G. Jørg., Fragilariopsis cylindrus (Grunow) Willi Krieg., Fragilariopsis sublinearis (Van Heurck) Heiden et Kolbe, C. closterium, Nitzschia lecointei Van Heurck, and the dinoflagellate Polarella glacialis Montresor, Procaccini et Stoecker. Two Arctic isolates were also examined, Nitzschia frigida (Grunow) and Fragilariopsis oceanica (Cleve) Hasle. Significant heterogeneity of protein expression was observed despite all cultures being grown in iron replete f ⁄ 2 medium. Only one species, F. cylindrus, displayed the expected expression of ferredoxin only in iron-replete medium. Four were observed to produce both proteins under iron-replete conditions. Ferredoxin was not detected at all in F. curta and Pseudo-nitzschia sp., but distinct flavodoxin bands were observed in both of these organisms. All species examined were observed to express either flavodoxin or ferredoxin or both of the proteins as determined by Western immunoblotting. Article in Journal/Newspaper Antarc* Antarctic Arctic ice algae Phytoplankton Sea ice Southern Ocean University of Tasmania: UTas ePrints Arctic Antarctic Southern Ocean The Antarctic Journal of Phycology 45 3 771 783 |
institution |
Open Polar |
collection |
University of Tasmania: UTas ePrints |
op_collection_id |
ftunivtasmania |
language |
English |
topic |
ferredoxin flavodoxin immunoassay; iron; microalgae; phytoplankton |
spellingShingle |
ferredoxin flavodoxin immunoassay; iron; microalgae; phytoplankton Pankowski, A McMinn, A Development of immunoassays for the iron-regulated proteins ferredoxin and flavodoxin in polar microalgae |
topic_facet |
ferredoxin flavodoxin immunoassay; iron; microalgae; phytoplankton |
description |
While the growth of Southern Ocean phytoplankton is often limited by iron availability, there are no comparable experiments on sea-ice algae. Here we assess the use of ferredoxin and flavodoxin to investigate the iron nutritional status of sea-ice algae and describe the development of a quantitative immunoassay for both proteins in marine diatoms. Highaffinity monoclonal antibodies toward both proteins were produced from Cylindrotheca closterium (Ehrenb.) J. M. Lewin et Reimann, and these were used to develop Western blots. Western blots run on whole protein extracts detected both proteins with little cross-reactivity toward other proteins. The two proteins could be successfully quantitated when applied to gels at between 5 and 50 ng in a volume of 25 lL (0.2–2 lg ÆmL)1). Flavodoxin and ferrodoxin expression was examined in the Antarctic diatoms Entomoneis kjellmannii (Cleve) Poulin et Cardinal, Navicula directa (W. Sm.) Ralfs, Fragilariopsis curta (Van Heurck) Hust., Pseudo-nitzschia sp., Porosira glacialis (Grunow) E. G. Jørg., Fragilariopsis cylindrus (Grunow) Willi Krieg., Fragilariopsis sublinearis (Van Heurck) Heiden et Kolbe, C. closterium, Nitzschia lecointei Van Heurck, and the dinoflagellate Polarella glacialis Montresor, Procaccini et Stoecker. Two Arctic isolates were also examined, Nitzschia frigida (Grunow) and Fragilariopsis oceanica (Cleve) Hasle. Significant heterogeneity of protein expression was observed despite all cultures being grown in iron replete f ⁄ 2 medium. Only one species, F. cylindrus, displayed the expected expression of ferredoxin only in iron-replete medium. Four were observed to produce both proteins under iron-replete conditions. Ferredoxin was not detected at all in F. curta and Pseudo-nitzschia sp., but distinct flavodoxin bands were observed in both of these organisms. All species examined were observed to express either flavodoxin or ferredoxin or both of the proteins as determined by Western immunoblotting. |
format |
Article in Journal/Newspaper |
author |
Pankowski, A McMinn, A |
author_facet |
Pankowski, A McMinn, A |
author_sort |
Pankowski, A |
title |
Development of immunoassays for the iron-regulated proteins ferredoxin and flavodoxin in polar microalgae |
title_short |
Development of immunoassays for the iron-regulated proteins ferredoxin and flavodoxin in polar microalgae |
title_full |
Development of immunoassays for the iron-regulated proteins ferredoxin and flavodoxin in polar microalgae |
title_fullStr |
Development of immunoassays for the iron-regulated proteins ferredoxin and flavodoxin in polar microalgae |
title_full_unstemmed |
Development of immunoassays for the iron-regulated proteins ferredoxin and flavodoxin in polar microalgae |
title_sort |
development of immunoassays for the iron-regulated proteins ferredoxin and flavodoxin in polar microalgae |
publishDate |
2009 |
url |
https://eprints.utas.edu.au/12240/ https://eprints.utas.edu.au/12240/1/DEVELOPMENT_OF_IMMUNOASSAYS_FOR_THE_IRON-REGULATED_PROTEINS_FERREDOXIN_AND_FLAVODOXIN_IN_POLAR_MICROALGAE1.pdf https://doi.org/10.1111/j.1529-8817.2009.00687.x |
geographic |
Arctic Antarctic Southern Ocean The Antarctic |
geographic_facet |
Arctic Antarctic Southern Ocean The Antarctic |
genre |
Antarc* Antarctic Arctic ice algae Phytoplankton Sea ice Southern Ocean |
genre_facet |
Antarc* Antarctic Arctic ice algae Phytoplankton Sea ice Southern Ocean |
op_relation |
https://eprints.utas.edu.au/12240/1/DEVELOPMENT_OF_IMMUNOASSAYS_FOR_THE_IRON-REGULATED_PROTEINS_FERREDOXIN_AND_FLAVODOXIN_IN_POLAR_MICROALGAE1.pdf Pankowski, A and McMinn, A 2009 , 'Development of immunoassays for the iron-regulated proteins ferredoxin and flavodoxin in polar microalgae' , Journal of Phycology, vol. 45 , pp. 771-783 , doi:10.1111/j.1529-8817.2009.00687.x <http://dx.doi.org/10.1111/j.1529-8817.2009.00687.x>. |
op_rights |
cc_utas |
op_doi |
https://doi.org/10.1111/j.1529-8817.2009.00687.x |
container_title |
Journal of Phycology |
container_volume |
45 |
container_issue |
3 |
container_start_page |
771 |
op_container_end_page |
783 |
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1766084033781956608 |