FtsH1 + FtsH5 | ATP-dependent zinc metalloprotease FtsH1 + FtsH5 (chloroplastic)
AS16 3930 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana, Nicotiana tabacum, Spinacia oleracea
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67.1 kD (Arabidopsis thaliana)
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Total proteins were isolated from Arabidopsis (Arabidopsis thaliana) wild type (Col) and mutant lacking FtsH2 (yellow variegated2 [var2]). Samples were immediately frozen in liquid nitrogen and pulverized with a microtube homogenizer. Proteins were extracted by adding appropriate extraction buffer. After measurement of chlorophyll concentration, equally loaded supernatants (based on chlorophyll [0.5 µg chlorophyll/lane]) were subjected to SDS-PAGE analysis. Proteins were separated on 12% SDS-PAGE gel and blotted 1h to PVDF membrane. Blots were blocked in 1% BSA in PBST buffer for 1 h at room temperature. Then, blots were incubated in the primary antibody (anti-VAR1) at a dilution of 1:5000 for 1 h. After washing 2 times for 10 min in PBST buffer, blots were incubated in the secondary antibody (anti Rabbit IgG) at a dilution of 1:5000 for 1 h. Blots were washed 2 times for 10 min in PBST buffer. Chemiluminescent detection reagent was used for signal detection. Images of the blots were obtained using ChemiDoc™ XRS (Bio-rad). Exposure time was 2 seconds.
Absence of FtsH2 in var2 mutant results in decreased amount of FtsH1 which together form a hetero-hexamer complex.
Courtesy of Dr. Yusuke Kato, Plant Light Acclimation Research Group, Okayama University, Japan
Both FtsH5 (VAR1) and FtsH1 ahare high degree of homology therefore this antibody recognizes both proteins.
FtsH belong to a family of ATP dependent peptidases. Localized in a chloroplast are following isoforms: FTSH1 (synonymes AAA, FTSH, FTSH Protease 1), Ftsh2 (VAR2, VARIEGATED 2), FtsH5 (VAR1, VARIEGATED 1), FtsH6 (FTSH PROTEASE 6), FtsH7, FtsH8. FtsH9. Localized in mitochondria are following isoforms: FtsH3, FtsH4, FtsH10, FtsH11.
FtsH5 (VAR1) is a component of the ATP-dependent zinc metallopeptidase. It is involved in the thylakoids biogenesis and in the repair of damaged D1 subunit of photosystem II, a process that protects against cell death under high light conditions. It forms a complex with VAR1. FtsH5 (VAR1) and FtsH1 are interchangeable in thylakoid membranes.