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Literature summary for 1.14.13.122 extracted from

  • Sakuraba, Y.; Tanaka, R.; Yamasato, A.; Tanaka, A.
    Determination of a chloroplast degron in the regulatory domain of chlorophyllide a oxygenase (2009), J. Biol. Chem., 284, 36689-36699.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 1.14.13.122

Cloned(Commentary)

Cloned (Comment) Organism
plasmid vector pGreenII-0029 including the gene for the green fluorescent protein fused with full-length or truncated chlorophyllide a oxygenase, plasmid with fusion enzyme is then transformed into Agrobacterium tumefaciens (strain GV2260) by electroporation, wild-type Arabidopsis and chlorina1-1 mutant are transformed by vacuum infiltration Arabidopsis thaliana

Protein Variants

Protein Variants Comment Organism
E136V amino acid mutation within A-domain, increased stability of the fusion enzyme, K157R alone without stability effect, deletions from Q97 up to or beyond H106 result in stability signal equivalent to a lack of the entire A-domain Arabidopsis thaliana
additional information serial deletions in the A-domain identify the amino acid sequence Q97-DLLTIMIL-H106 that is essential for the regulation of the protein stability, this sequence introduced into mutant chlorina1-1 fusion enzyme lacking the entire A-domain re-enables the regulation of the enzyme, the other part of the A-domain is necessary to confer chlrorophyll-b dependency of the degradation process 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 chlorophyll a precursor chlorophyllide b + 2 H2O + NADP+ immediately converted to chlorophyll b by chlorophyll synthase, the chlorophyllide a oxygenase activity is regulated at the level of protein stability via a feedback mechanism through chlorophyll b ?
additional information Arabidopsis thaliana N-terminal domain (A-domain) of enzyme is essential for the regulatory mechanism of the enzyme ?
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 ?

Organism

Organism UniProt Comment Textmining
Arabidopsis thaliana
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 wild ecotype Columbia and chlorina1-1 mutant (lacking the chlorophyllide a oxigenase)
-

Purification (Commentary)

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

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
chlorophyllide a + O2 + NADPH + H+ chlorophyll a precursor Arabidopsis thaliana chlorophyllide b + 2 H2O + NADP+ immediately converted to chlorophyll b by chlorophyll synthase, the chlorophyllide a oxygenase activity is regulated at the level of protein stability via a feedback mechanism through chlorophyll b ?
additional information N-terminal domain (A-domain) of enzyme is essential for the regulatory mechanism of the enzyme Arabidopsis thaliana ?
-
 ?

Synonyms

Synonyms Comment Organism
CAO
-
 Arabidopsis thaliana
chlorophyllide a oxygenase
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 Arabidopsis thaliana