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Literature summary for 4.99.1.3 extracted from

  • Lobo, S.A.; Brindley, A.A.; Romao, C.V.; Leech, H.K.; Warren, M.J.; Saraiva, L.M.
    Two distinct roles for two functional cobaltochelatases (CbiK) in Desulfovibrio vulgaris Hildenborough (2008), Biochemistry, 47, 5851-5857.
    View publication on PubMed
Search for more literature for 4.99.1.3

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Desulfovibrio vulgaris
gene DVU0650, DNA and amino acid sequence determination and analysis, expression of wild-type enzyme and mutant DELTA28DVU0650 in Escherichia coli as His-tagged proteins, complementation of Escherichia coli cysG mutant strain by DELTA28CbiKP Desulfovibrio vulgaris
gene DVU1365, DNA and amino acid sequence determination and analysis, expression of wild-type enzyme in Escherichia coli as His-tagged protein, complementation of Escherichia coli cysG mutant strain by CbiKC Desulfovibrio vulgaris

Protein Variants

Protein Variants Comment Organism
additional information CbiKc gene is able to complement an Escherichia coli cysG mutant Desulfovibrio vulgaris
additional information CbiKp gene is able to complement an Escherichia coli cysG mutant Desulfovibrio vulgaris
additional information construction of a truncated DVU0650 protein that starts at amino acid 29, resulting in mutant DELTA28DVU0650. A truncated protein that lacks the signal peptide is also produced, named DELTA28CbiKP Desulfovibrio vulgaris

Localization

Localization Comment Organism GeneOntology No. Textmining
cytoplasm enzyme may be involved in cytoplasmic cobalamin biosynthesis Desulfovibrio vulgaris 5737
-
cytoplasm CbiKc, the isozyme lacks an N-terminal transit peptide Desulfovibrio vulgaris 5737
-
periplasm
-
 Desulfovibrio vulgaris
-
-
periplasm CbiKp Desulfovibrio vulgaris
-
-

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
28000
-
 x * 28000, SDS-PAGE and calculated, mature protein Desulfovibrio vulgaris
31000
-
 x * 31000, recombinant enzyme, SDS-PAGE Desulfovibrio vulgaris
31000
-
 x * 31000, PAGE and calculated, mature protein Desulfovibrio vulgaris

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
additional information Desulfovibrio vulgaris CbiKC is likely to be the enzyme associated with cytoplasmic cobalamin biosynthesis ?
-
 ?
additional information Desulfovibrio vulgaris CbiKP is periplasmic located and possibly associated with an iron transport system ?
-
 ?
additional information Desulfovibrio vulgaris Hildenborough CbiKC is likely to be the enzyme associated with cytoplasmic cobalamin biosynthesis ?
-
 ?
additional information Desulfovibrio vulgaris Hildenborough CbiKP is periplasmic located and possibly associated with an iron transport system ?
-
 ?
sirohydrochlorin + Co2+ Desulfovibrio vulgaris a step involved in the vitamin B12 biosynthesis cobalt-sirohydrochlorin + 2 H+
-
 ?
sirohydrochlorin + Co2+ Desulfovibrio vulgaris Hildenborough a step involved in the vitamin B12 biosynthesis cobalt-sirohydrochlorin + 2 H+
-
 ?
sirohydrochlorin + Fe2+ Desulfovibrio vulgaris
-
 iron-sirohydrochlorin + 2 H+
-
 ?

Organism

Organism UniProt Comment Textmining
Desulfovibrio vulgaris Q72CB8 isoform CbiKc
-
Desulfovibrio vulgaris Q72CB8 DVU_1365
-
Desulfovibrio vulgaris Q72EC8 isoform CbiKp
-
Desulfovibrio vulgaris Q72EC8 DVU_0650
-
Desulfovibrio vulgaris Hildenborough Q72CB8 isoform CbiKc
-
Desulfovibrio vulgaris Hildenborough Q72EC8 isoform CbiKp
-

Posttranslational Modification

Posttranslational Modification Comment Organism
additional information enzyme lacks a signal peptide Desulfovibrio vulgaris
proteolytic modification processing of 28 amino acid signal peptide Desulfovibrio vulgaris

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged wild-type isozyme and mutant DELTA28DVU0650 from Escherichia coli by nickel affinity chromatography, dialysis, anion exchange chromatography and gel filtration Desulfovibrio vulgaris
recombinant His-tagged wild-type isozyme from Escherichia coli by nickel affinity chromatography, dialysis, anion exchange chromatography and gel filtration Desulfovibrio vulgaris

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.0015
-
 purified recombinant CbiKc, with Fe2+ Desulfovibrio vulgaris
0.004
-
 purified recombinant CbiKc, with Co2+ Desulfovibrio vulgaris
1.5
-
 substrate Fe2+, pH 8.0 Desulfovibrio vulgaris
4
-
 substrate Co2+, pH 8.0 Desulfovibrio vulgaris
13
-
 substrate Fe2+, pH 8.0 Desulfovibrio vulgaris
22
-
 substrate Co2+, pH 8.0 Desulfovibrio vulgaris

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + sirohydrochlorin + Co2+
-
 Desulfovibrio vulgaris ADP + phosphate + Co(II)-sirohydrochlorin + H+
-
 ?
ATP + sirohydrochlorin + Co2+
-
 Desulfovibrio vulgaris Hildenborough ADP + phosphate + Co(II)-sirohydrochlorin + H+
-
 ?
ATP + sirohydrochlorin + Fe2+
-
 Desulfovibrio vulgaris ADP + phosphate + Fe(II)-sirohydrochlorin + H+
-
 ?
ATP + sirohydrochlorin + Fe2+
-
 Desulfovibrio vulgaris Hildenborough ADP + phosphate + Fe(II)-sirohydrochlorin + H+
-
 ?
additional information CbiKC is likely to be the enzyme associated with cytoplasmic cobalamin biosynthesis Desulfovibrio vulgaris ?
-
 ?
additional information CbiKP is periplasmic located and possibly associated with an iron transport system Desulfovibrio vulgaris ?
-
 ?
additional information CbiKC is likely to be the enzyme associated with cytoplasmic cobalamin biosynthesis Desulfovibrio vulgaris Hildenborough ?
-
 ?
additional information CbiKP is periplasmic located and possibly associated with an iron transport system Desulfovibrio vulgaris Hildenborough ?
-
 ?
sirohydrochlorin + Co2+ a step involved in the vitamin B12 biosynthesis Desulfovibrio vulgaris cobalt-sirohydrochlorin + 2 H+
-
 ?
sirohydrochlorin + Co2+ both CbiKP and DELTA28CbiKP insert cobalt and iron into sirohydrochlorin with specific activity with iron lower than that measured with cobalt Desulfovibrio vulgaris cobalt-sirohydrochlorin + 2 H+
-
 ?
sirohydrochlorin + Co2+ the enzyme inserts cobalt and iron into sirohydrochlorin with specific activity with iron lower than that measured with cobalt Desulfovibrio vulgaris cobalt-sirohydrochlorin + 2 H+
-
 ?
sirohydrochlorin + Co2+ a step involved in the vitamin B12 biosynthesis Desulfovibrio vulgaris Hildenborough cobalt-sirohydrochlorin + 2 H+
-
 ?
sirohydrochlorin + Co2+ the enzyme inserts cobalt and iron into sirohydrochlorin with specific activity with iron lower than that measured with cobalt Desulfovibrio vulgaris Hildenborough cobalt-sirohydrochlorin + 2 H+
-
 ?
sirohydrochlorin + Co2+ both CbiKP and DELTA28CbiKP insert cobalt and iron into sirohydrochlorin with specific activity with iron lower than that measured with cobalt Desulfovibrio vulgaris Hildenborough cobalt-sirohydrochlorin + 2 H+
-
 ?
sirohydrochlorin + Fe2+
-
 Desulfovibrio vulgaris iron-sirohydrochlorin + 2 H+
-
 ?
sirohydrochlorin + Fe2+ both CbiKP and DELTA28CbiKP insert cobalt and iron into sirohydrochlorin with specific activity with iron lower than that measured with cobalt Desulfovibrio vulgaris iron-sirohydrochlorin + 2 H+
-
 ?
sirohydrochlorin + Fe2+ the enzyme inserts cobalt and iron into sirohydrochlorin with specific activity with iron lower than that measured with cobalt Desulfovibrio vulgaris iron-sirohydrochlorin + 2 H+
-
 ?

Subunits

Subunits Comment Organism
? x * 31000, recombinant enzyme, SDS-PAGE Desulfovibrio vulgaris
? x * 28000, SDS-PAGE and calculated, mature protein Desulfovibrio vulgaris
? x * 31000, PAGE and calculated, mature protein Desulfovibrio vulgaris

Synonyms

Synonyms Comment Organism
CbiK
-
 Desulfovibrio vulgaris
CbiKc
-
 Desulfovibrio vulgaris
CbiKp
-
 Desulfovibrio vulgaris
cobaltochelatase
-
 Desulfovibrio vulgaris

Cofactor

Cofactor Comment Organism Structure
heme heme b prosthetic group Desulfovibrio vulgaris