Information on EC 6.6.1.2 - cobaltochelatase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
6.6.1.2
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RECOMMENDED NAME
GeneOntology No.
cobaltochelatase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
ATP + hydrogenobyrinic acid a,c-diamide + Co2+ + H2O = ADP + phosphate + cob(II)yrinic acid a,c-diamide + H+
show the reaction diagram
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
cob(II)yrinate a,c-diamide biosynthesis II (late cobalt incorporation)
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Metabolic pathways
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Porphyrin and chlorophyll metabolism
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vitamin B12 metabolism
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SYSTEMATIC NAME
IUBMB Comments
hydrogenobyrinic-acid-a,c-diamide:cobalt cobalt-ligase (ADP-forming)
This enzyme, which forms part of the aerobic cobalamin biosynthesis pathway, is a type I chelatase, being heterotrimeric and ATP-dependent. It comprises two components, one of which corresponds to CobN and the other is composed of two polypeptides, specified by cobS and cobT in Pseudomonas denitrificans, and named CobST [1]. Hydrogenobyrinic acid is a very poor substrate. ATP can be replaced by dATP or CTP but the reaction proceeds more slowly. CobN exhibits a high affinity for hydrogenobyrinic acid a,c-diamide. The oligomeric protein CobST possesses at least one sulfhydryl group that is essential for ATP-binding. Once the Co2+ is inserted, the next step in the pathway ensures that the cobalt is ligated securely by reducing Co(II) to Co(I). This step is carried out by EC 1.16.8.1, cob(II)yrinic acid a,c-diamide reductase.
CAS REGISTRY NUMBER
COMMENTARY hide
81295-49-0
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
cobaltochelatase consists of three subunits, CobN, CobS and CobT, while subunits CobS and CobT form a chaperone-like complex, characteristic for the AAA+ class of proteins
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Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
ATP + hydrogenobyrinic acid a,c-diamide + Co2+
ADP + phosphate + cob(II)yrinic acid a,c-diamide + H+
show the reaction diagram
CTP + hydrogenobyrinic acid a,c-diamide + Co2+
CDP + phosphate + cob(II)yrinic acid a,c-diamide + H+
show the reaction diagram
P29933 AND P29934 AND Q9HZQ3 AND P29929
45% of activity with respect to ATP
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?
dATP + hydrogenobyrinic acid a,c-diamide + Co2+
dADP + phosphate + cob(II)yrinic acid a,c-diamide + H+
show the reaction diagram
P29933 AND P29934 AND Q9HZQ3 AND P29929
65% of activity with respect to ATP
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?
hydrogenobyrinic acid a,c-diamide + ATP + Co2+
ADP + phosphate + cob(II)yrinic acid a,c-diamide + H+
show the reaction diagram
P29933 AND P29934 AND Q9HZQ3 AND P29929
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?
ITP + hydrogenobyrinic acid a,c-diamide + Co2+
IDP + phosphate + cob(II)yrinic acid a,c-diamide + H+
show the reaction diagram
P29933 AND P29934 AND Q9HZQ3 AND P29929
5% of activity with respect to ATP
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?
additional information
?
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the protein binds cobalt via residues His154, Glu184, and His216
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NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
ATP + hydrogenobyrinic acid a,c-diamide + Co2+
ADP + phosphate + cob(II)yrinic acid a,c-diamide + H+
show the reaction diagram
hydrogenobyrinic acid a,c-diamide + ATP + Co2+
ADP + phosphate + cob(II)yrinic acid a,c-diamide + H+
show the reaction diagram
P29933 AND P29934 AND Q9HZQ3 AND P29929
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?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
heme b
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Mg2+
P29933 AND P29934 AND Q9HZQ3 AND P29929
ATP binding cation
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
adenylyl (beta,gamma-methylene)-diphosphonate
P29933 AND P29934 AND Q9HZQ3 AND P29929
50% inhibition at 5 mM
adenylyl-imidodiphosphate
P29933 AND P29934 AND Q9HZQ3 AND P29929
50% inhibition at 2 mM
ADP
P29933 AND P29934 AND Q9HZQ3 AND P29929
50% inhibition at 8 mM
AMP
P29933 AND P29934 AND Q9HZQ3 AND P29929
15% inhibition at 10 mM
Cu2+
P29933 AND P29934 AND Q9HZQ3 AND P29929
50% inhibition at 0.002 mM
diphosphate
P29933 AND P29934 AND Q9HZQ3 AND P29929
50% inhibition at 5 mM
Fe2+
P29933 AND P29934 AND Q9HZQ3 AND P29929
50% inhibition at 0.05 mM
N-ethylmaleimide
P29933 AND P29934 AND Q9HZQ3 AND P29929
50% inhibition at 0.005 mM
Ni2+
P29933 AND P29934 AND Q9HZQ3 AND P29929
50% inhibition at 0.003 mM
Zn2+
P29933 AND P29934 AND Q9HZQ3 AND P29929
50% inhibition at 0.0005 mM
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.22
ATP
P29933 AND P29934 AND Q9HZQ3 AND P29929
pH 8.0, 30°C
0.0042
Co2+
P29933 AND P29934 AND Q9HZQ3 AND P29929
pH 8.0, 30°C
0.000085
hydrogenobyrinic acid a,c-diamide
P29933 AND P29934 AND Q9HZQ3 AND P29929
pH 8.0, 30°C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
38000
P29933 AND P29934 AND Q9HZQ3 AND P29929
1 * 140000, CobN + 1 * 38000, namely CobS + 1 * 80000, namely CobT, gel filtration, SDS-PAGE
80000
P29933 AND P29934 AND Q9HZQ3 AND P29929
1 * 140000, CobN + 1 * 38000, namely CobS + 1 * 80000, namely CobT, gel filtration, SDS-PAGE
140000
P29933 AND P29934 AND Q9HZQ3 AND P29929
1 * 140000, CobN + 1 * 38000, namely CobS + 1 * 80000, namely CobT, gel filtration, SDS-PAGE
258000
P29933 AND P29934 AND Q9HZQ3 AND P29929
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
tetramer
trimer
P29933 AND P29934 AND Q9HZQ3 AND P29929
1 * 140000, CobN + 1 * 38000, namely CobS + 1 * 80000, namely CobT, gel filtration, SDS-PAGE
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
hanging drop vapor diffusion method, using 100 mM Tris-HCl. pH 8.5, containing 2.0 M ammonium sulfate
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
purification includes: Mono Q HR 10/10, Penyl-Superose and Mono Q HR 5/5 chromatographies for CobN subunit, Mono Q HR 10/10, Phenyl-Superose and AGATP for CobST subunit
P29933 AND P29934 AND Q9HZQ3 AND P29929
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
subunits separately overexpressed in Escherichia coli BL21star (DE3)pLysS
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