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Development of Immunoassays for the Iron-Regulated Proteins Ferredoxin and Flavodoxin in Polar Microalgae
Citation
Pankowski, A and McMinn, A, Development of Immunoassays for the Iron-Regulated Proteins Ferredoxin and Flavodoxin in Polar Microalgae, Journal of Phycology, 45, (3) pp. 771-783. ISSN 0022-3646 (2009) [Refereed Article]
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Copyright Statement
The definitive published version is available online at: http://interscience.wiley.com
Official URL: http://interscience.wiley.com
DOI: doi:10.1111/j.1529-8817.2009.00687.x
Abstract
While the growth of Southern Ocean phytoplank-
ton 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 inves-
tigate the iron nutritional status of sea-ice algae and
describe the development of a quantitative immuno-
assay for both proteins in marine diatoms. High-
affinity 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 Æ mL)1). Flavodoxin and ferrodox-
in 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 gla-
cialis (Grunow) E. G. Jørg., Fragilariopsis cylindrus
(Grunow) Willi Krieg., Fragilariopsis sublinearis (Van
Heurck) Heiden et Kolbe, C. closterium, Nitzschia leco-
intei Van Heurck, and the dinoflagellate Polarella gla-
cialis Montresor, Procaccini et Stoecker. Two Arctic
isolates were also examined, Nitzschia frigida (Gru-
now) and Fragilariopsis oceanica (Cleve) Hasle. Signif-
icant 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.
Item Details
Item Type: | Refereed Article |
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Keywords: | ferredoxin, flavodoxin, immunoassay, iron, microalgae, phytoplankton |
Research Division: | Biological Sciences |
Research Group: | Ecology |
Research Field: | Marine and estuarine ecology (incl. marine ichthyology) |
Objective Division: | Environmental Management |
Objective Group: | Marine systems and management |
Objective Field: | Marine biodiversity |
UTAS Author: | Pankowski, A (Mr Andrew Pankowski) |
UTAS Author: | McMinn, A (Professor Andrew McMinn) |
ID Code: | 57404 |
Year Published: | 2009 |
Web of Science® Times Cited: | 16 |
Deposited By: | IASOS |
Deposited On: | 2009-07-16 |
Last Modified: | 2010-05-07 |
Downloads: | 0 |
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