PsbA | D1 protein of PSII, C-terminal (chicken)
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KLH-conjugated synthetic peptide derived from available plant, algal and cyanobacterial PsbA sequences, including Arabidopsis thaliana UniProt: A4QJR4, TAIR: AtCg00020 , Oryza sativa P0C434, Populus alba Q14FH6, Physcomitrella patens Q6YXN7, Chlamydomonas reinhardtii P07753, Synechocystis sp. P14660 and many others
38 | 28-30 kDa
Algae (brown and red), Conifers, Cryptomonads, Legumes, Stramenopiles, Euglenoids, Prochlorophytes, Xantophytes
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2 µg of total protein from (1) Arabidopsis thaliana leaf extracted with PEB (AS08 300), (2) Horderum vulgare leaf extracted with PEB (AS08 300), (3) Chlamydomonas reinhardtii total cell extracted with PEB (AS08 300), (4) Synechococcus sp. 7942 total cell extracted with PEB (AS08 300), (5) Anabaena sp. total cell extracted with PEB (AS08 300) were separated on 4-12% NuPage (Invitrogen) LDS-PAGE and blotted 1h to PVDF. Blots were blocked immediately following transfer for 1h at room temperature with agitation. Blots were incubated in the primary antibody at a dilution of 1: 50 000 for 1h at room temperature with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at room temperature with agitation. Blots were incubated in secondary antibody (anti-hen IgY horse radish peroxidase conjugated, recommended secondary antibody AS09 603) diluted to 1:50 000 for 1h/RT with agitation. The blots were washed as above and developed for 5 min with chemiluminescent detection reagent, according the manufacturers instructions. Images of the blots were obtained using a CCD imager (FluorSMax, Bio-Rad) and Quantity One software (Bio-Rad).
Reactant: Arabidopsis thaliana (Thale cress)
Application: Western Blotting
Pudmed ID: 28791032
Journal: Front Plant Sci
Figure Number: 2A
Published Date: 2017-08-10
First Author: Kohzuma, K., Froehlich, J. E., et al.
Impact Factor: 5.435Open Publication
Changes in the protein levels of photosynthetic components under extended dark exposure in wild-type and gamera-1. Immunoblot detection of photosynthetic proteins from leaves of Ws and gamera-1 plants incubated after dark adaptation for 0, 2, and 4 days was examined. Specifically, essentially thylakoid fractions were assayed to determine the content of the following proteins: ?-subunit of ATP synthase; the D1 protein, OEC17, OEC23, and OEC33 of PSII; Cyt f and Rieske protein of the cytochrome b6f complex; and the F and D subunits of PSI, after extended dark treatment. Proteins were resolved via SDS-PAGE gel based on equal microgram chlorophyll per lane loading and processed as described in Section “Materials and Methods”. The Large subunit of RuBisco and LHCII stained with either CBB or Ponceau red, respectively, are presented here as loading controls. DAD indicates days after dark adaptation.
A number of degradation products may be observed when using anti-PsbA antibodies, including products having apparent molecular weights of 24kDa and 16kDa. D1 degradation is a complex set of events and the products observed can be influenced by both the extraction procedure and the physiology of the cells prior to harvest. Third, cross-linking may occur between D1 and cytochrome b559, shifting the protein higher in the gel. In cyanobacteria (PCC7942), three different bands were competed out by preincubating the antibody with the PsbA free peptide, indicating that all bands are indeed PsbA and its precursors or breakdown products. Competition assays were also performed with spinach and Chlamydomonas, confirming the identity of PsbA bands. Anti-PsbA antibodies will not detect D2 protein, as the peptide used to generate PsbA antibodies has no homology to the D2 sequence.
Example of a simulataneous western blot detection with RbcL, PsbA and PsaC antibodies.
The antibody is appropriate for detecting both, 24 kDa or the 10 kDa C-terminal fragments, whichever is generated under given treatment conditions.In our analysis we have seen both, ca. 24 kDa and ca. 10 kDa fragments from different samples, depending on treatments and isolation procedures.
This antibody will also detect the phosphorylated form of D1as an alternate band to the main band on a high resolution gel.
The PsbA (D1) protein of Photosystem II is rapidly cycled under illumination in all oxygenic photobionts. Disruption of PsbA cycling or losses of PsbA pools are central to photoinhibition of photosynthesis in cyanobacteria, algae and plants under a wide range of conditions including excess light, low temperature and UV exposure. Tracking PsbA pools using the Global PsbA antibody can show the functional content of Photosystem II in a wide range of samples.
Sicora et al. (2019). Regulation of PSII function in Cyanothece sp. ATCC 51142 during a light-dark cycle. Photosynth Res. 2019 Mar;139(1-3):461-473. doi: 10.1007/s11120-018-0598-5,
Sevilla et al. (2019). Regulation by FurC in Anabaena links the oxidative stress response to photosynthetic metabolism. Plant Cell Physiol. 2019 May 21. pii: pcz094. doi: 10.1093/pcp/pcz094.
Figlioli et al. (2019). Overall plant responses to Cd and Pb metal stress in maize: Growth pattern, ultrastructure, and photosynthetic activity. Environ Sci Pollut Res Int. 2019 Jan;26(2):1781-1790. doi: 10.1007/s11356-018-3743-y.
Krupinska et al. (2019). The nucleoid-associated protein WHIRLY1 is required for the coordinate assembly of plastid and nucleus-encoded proteins during chloroplast development. Planta. 2019 Jan 11. doi: 10.1007/s00425-018-03085-z.
Sicora et al. (2018). Regulation of PSII function in Cyanothece sp. ATCC 51142 during a light–dark cycle. Photosynth Res. 2018 Oct 24. doi: 10.1007/s11120-018-0598-5.
Lentini et al. (2018). Early responses to cadmium exposure in barley plants: effects on biometric and physiological parameters. Acta Physiol Plant (2018) 40: 178. https://doi.org/10.1007/s11738-018-2752-2.
Gonzaga Heredia-Martinez et al. (2018). Chloroplast damage induced by the inhibition of fatty acid synthesis triggers autophagy in Chlamydomonas. Plant Physiol, Sept. 2018.
Kong et al. (2018) Interorganelle Communication: Peroxisomal MALATE DEHYDROGENASE2 Connects Lipid Catabolism to Photosynthesis through Redox Coupling in Chlamydomonas. Plant Cell. 2018 Aug;30(8):1824-1847. doi: 10.1105/tpc.18.00361
Dy et al. (2018). Galactoglycerolipid Lipase PGD1 Is Involved in Thylakoid Membrane Remodeling in Response to Adverse Environmental Conditions in Chlamydomonas. Plant Cell. 2018 Feb;30(2):447-465. doi: 10.1105/tpc.17.00446.
Kim et al. (2018). The rice zebra3 (z3) mutation disrupts citrate distribution and produces transverse dark-green/green variegation in mature leaves. Rice (N Y). 2018 Jan 5;11(1):1. doi: 10.1186/s12284-017-0196-8.
Yokono et al. (2015). A megacomplex composed of both photosystem reaction centres in higher plants. Nat Commun. 2015 Mar 26;6:6675. doi: 10.1038/ncomms7675.
Su et al. (2014). Exogenous progesterone alleviates heat and high light stress-induced inactivation of photosystem II in wheat by enhancing antioxidant defense and D1 protein stability. Plant Growth Regul DOI 0.1007/s10725-014-9920-1
Esparza et al. (2013). Katanin Localization Requires Triplet Microtubules in Chlamydomonas reinhardtii. PLOS one.
Hoogenboom et al. (2012). Effects of Light, Food Availability and Temperature Stress on the Function of Photosystem II and Photosystem I of Coral Symbionts. POLS one.
Morash et al. (2007). Macromolecular dynamics of the photosynthetic system over a seasonal developmental progression in Spartina alterniflora. Canadian J. of Botany, 2007, 85(5): 476-483, 10.1139/B07-043.
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