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Literature summary for 7.2.2.2 extracted from

  • Smith, A.T.; Ross, M.O.; Hoffman, B.M.; Rosenzweig, A.C.
    Metal selectivity of a Cd-, Co-, and Zn-transporting P1B-type ATPase (2017), Biochemistry, 56, 85-95.
    View publication on PubMed
Search for more literature for 7.2.2.2

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Cupriavidus metallidurans

Protein Variants

Protein Variants Comment Organism
C476A the mutant enzyme binds 0.61 mol equiv of Co2+. The mutant enzyme shows about 4%, about 24%, and about 30% of Cd2+-, Co2+-, and Zn2+-stimulated ATP hydrolysis activity, respectively, compared to the wild type enzyme Cupriavidus metallidurans
H807A the mutant enzyme retains the ability to bind about 1 mol equiv of all metal ions (Co2+, Zn2+, and Cd2+). The mutation results in a strong decrease in Co2+- (about 17% of wild type) and Zn2+-stimulated (about 16% of wild type) ATP hydrolysis Cupriavidus metallidurans
M254A the mutant enzyme binds 0.34 mol and 0.26 mol equiv of Cd2+ and Co2+, respectively. The mutation dramatically reduces Cd2+- and Co2+-stimulated ATPase activity (about 8% and about 32% of wild type activity, respectively), whereas Zn2+-stimulated ATPase activity remains virtually unchanged Cupriavidus metallidurans
S474A the mutant enzyme binds 0.45 mol equiv of Co2+. The mutation modestly decreases Co2+-stimulated ATP hydrolysis (about 66% of wild type activity) whereas Cd2+- and Zn2+-stimulated ATP hydrolysis remain virtually unchanged Cupriavidus metallidurans
S474A/C476A the mutant enzyme shows about 2%, about 16%, and about 20% of Cd2+-, Co2+-, and Zn2+-stimulated ATP hydrolysis activity, respectively, compared to the wild type enzyme Cupriavidus metallidurans

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ATP + H2O + Cd2+[side 1] Cupriavidus metallidurans highest activity ADP + phosphate + Cd2+[side 2]
-
 ?
ATP + H2O + Co2+[side 1] Cupriavidus metallidurans
-
 ADP + phosphate + Co2+[side 2]
-
 ?
ATP + H2O + Zn2+[side 1] Cupriavidus metallidurans lowest activity ADP + phosphate + Zn2+[side 2]
-
 ?

Organism

Organism UniProt Comment Textmining
Cupriavidus metallidurans Q1LAJ7 formerly named Alcaligenes eutrophus and Ralstonia metallidurans
-

Purification (Commentary)

Purification (Comment) Organism
Ni-charged HiTrap column chromatography and Ni-NTA column chromatography Cupriavidus metallidurans

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.04
-
 with Cd2+ as transport ion, at pH 7.5 and 30°C Cupriavidus metallidurans
0.06
-
 with Co2+ as transport ion, at pH 7.5 and 30°C Cupriavidus metallidurans
0.2
-
 with Cd2+ as transport ion, at pH 7.5 and 30°C Cupriavidus metallidurans

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + H2O + Cd2+[side 1] highest activity Cupriavidus metallidurans ADP + phosphate + Cd2+[side 2]
-
 ?
ATP + H2O + Co2+[side 1]
-
 Cupriavidus metallidurans ADP + phosphate + Co2+[side 2]
-
 ?
ATP + H2O + Zn2+[side 1] lowest activity Cupriavidus metallidurans ADP + phosphate + Zn2+[side 2]
-
 ?

Synonyms

Synonyms Comment Organism
CzcP
-
 Cupriavidus metallidurans