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Literature summary for 1.14.13.122 extracted from

  • Kanematsu, S.; Sakuraba, Y.; Tanaka, A.; Tanaka, R.
    Characterization of Arabidopsis mutants defective in the regulation of chlorophyllide a oxygenase (2008), Photochem. Photobiol. Sci., 7, 1196-1205.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
pGreenII-0029 plasmid vector with fusion enzyme is introduced into Agrobacterium tumefaciens strain C58 by electroporation, wild-type and chlorina1-1 mutants of Arabidopsis thaliana are transformed by the infiltration with the bacterium, plant transformants are selected Arabidopsis thaliana

Protein Variants

Protein Variants Comment Organism
additional information seeds overexpressing A+B-domain-green fluorescent protein (AB-GFP) fusion enzyme, mutagenized by ethyl methane sulfonate overnight, 30000 lines of mutagenized transgenic plants screened for GFP fluorescence (excitation at 488 nm, emission at 522 nm) and observation of autofluorescence of chlorophyll (680 nm), 66 mutants with GFP signal isolated and grouped into 7 groups that are defined by the distribution pattern of the GFP signal, and characterized by pigment and immunoblotting analysis Arabidopsis thaliana

Localization

Localization Comment Organism GeneOntology No. Textmining
chloroplast mutant fusion enzyme signal patterns in chloroplasts, fusion enzyme signal only in vacuolated chloroplasts (abnormally developed, no organized thylakoid membranes and chlorophylls) within variegated leaf sectors (group IV mutants - defect in chloroplast development) Arabidopsis thaliana 9507
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plasma membrane only fusion enzyme localization (group VII mutant - defect in cellular trafficking of chloroplast proteins) Arabidopsis thaliana 5886
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Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
48000
-
A and B domain + green fluorescence protein(AB-GFP) fusion enzyme, SDS-PAGE Arabidopsis thaliana
56000
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full-length enzyme, SDS-PAGE Arabidopsis thaliana

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
chlorophyllide a + O2 + NADPH + H+ Arabidopsis thaliana intermediate 7-hydroxymetyl(ide) chlorophyll a chlorophyllide b + H2O + NADP+
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?

Organism

Organism UniProt Comment Textmining
Arabidopsis thaliana
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wild-type, chlorina1-1 mutant (lacking the endogenous chlorophyllide a oxigenase), and clpC1-mutant (deficient in the subunit of the chloroplast Clp protease, acting as a molecular chaperone recognizing the protease substrate sequence), transgenic plants overexpressing a chimeric protein consisting of the A (destabilized enzyme in the presence of chlorophyll b) and B domains of the enzyme and green fluorescent protein (GFP), seeds mutagenized by ethyl methane sulfonate, screened for GFP fluorescence
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Purification (Commentary)

Purification (Comment) Organism
for immunoblotting analysis rosette leaves are homogenized with extraction buffer (50 mM Tris, pH 6.8, 2 mM EDTA, 10% glycerol, 2% SDS, 6% 2-mercaptoethanol), centrifugation, supernatant separated by SDS-PAGE, resolved proteins electroblotted onto Hybond-P membrane, identification of fusion enzyme signal with antibody-horseradish peroxidase system Arabidopsis thaliana

Source Tissue

Source Tissue Comment Organism Textmining
epidermal cell small young cells adjacent to the epidermis cells with less frequent fusion enzyme signal (group III mutants), fusion enzyme signals only in small young cells adjacent to the epidermis cells (group V mutants - impaired chloroplast development with less chlorophyll) Arabidopsis thaliana
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guard cell cotyledons, uniform distribution of the fusion enzyme signal within the chloroplast (group 1 mutants - defects in specific steps of the chlorophyllide a enzyme regulation), punctate fusion enzyme signal (group II mutants - defect in chloroplast proteases), fusion enzyme signal only in guard cells (group VI mutants - slight inhibition of enzyme degradation or slightly enhanced enzyme synthesis) Arabidopsis thaliana
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leaf chloroplasts of mesophyll and guard cells in cotyledons Arabidopsis thaliana
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mesophyll cotyledons, uniform distribution of the fusion enzyme signal within the chloroplast (group I mutants - defects in specific steps of the chlorophyllide a enzyme regulation), punctate fusion enzyme signal (group II mutants - defect in chloroplast proteases), fusion enzyme signal dispersed within chloroplasts and at the same time high-intensity signals localized in confined areas within chloroplasts (group III mutants) Arabidopsis thaliana
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Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
chlorophyllide a + O2 + NADPH + H+ intermediate 7-hydroxymetyl(ide) chlorophyll a Arabidopsis thaliana chlorophyllide b + H2O + NADP+
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?

Synonyms

Synonyms Comment Organism
CAO
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Arabidopsis thaliana
chlorophyllide a oxygenase
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Arabidopsis thaliana