Bacterial ferrochelatase turns human: Tyr13 determines the apparent metal specificity of Bacillus subtilis ferrochelatase.

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Bacterial ferrochelatase turns human: Tyr13 determines the apparent metal specificity of Bacillus subtilis ferrochelatase. / Hansson, Mattias; Karlberg, Tobias; Söderberg, Christopher; Rajan, Sreekanth; Warren, Martin J; Al-Karadaghi, Salam; Rigby, Stephen E J; Hansson, Mats.

I: Journal of Biological Inorganic Chemistry, Vol. 16, Nr. 2, 2011, s. 235-242.

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Hansson, Mattias ; Karlberg, Tobias ; Söderberg, Christopher ; Rajan, Sreekanth ; Warren, Martin J ; Al-Karadaghi, Salam ; Rigby, Stephen E J ; Hansson, Mats. / Bacterial ferrochelatase turns human: Tyr13 determines the apparent metal specificity of Bacillus subtilis ferrochelatase. I: Journal of Biological Inorganic Chemistry. 2011 ; Vol. 16, Nr. 2. s. 235-242.

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TY - JOUR

T1 - Bacterial ferrochelatase turns human: Tyr13 determines the apparent metal specificity of Bacillus subtilis ferrochelatase.

AU - Hansson, Mattias

AU - Karlberg, Tobias

AU - Söderberg, Christopher

AU - Rajan, Sreekanth

AU - Warren, Martin J

AU - Al-Karadaghi, Salam

AU - Rigby, Stephen E J

AU - Hansson, Mats

PY - 2011

Y1 - 2011

N2 - Ferrochelatase catalyzes the insertion of Fe(2+) into protoporphyrin IX. The enzymatic product heme (protoheme IX) is a well-known cofactor in a wide range of proteins. The insertion of metal ions other than Fe(2+) occurs rarely in vivo, but all ferrochelatases that have been studied can insert Zn(2+) at a good rate in vitro. Co(2+), but not Cu(2+), is known to be a good substrate of the mammalian and Saccharomyces cerevisiae ferrochelatases. In contrast, Cu(2+), but not Co(2+), has been found to be a good substrate of bacterial Bacillus subtilis ferrochelatase. It is not known how ferrochelatase discriminates between different metal ion substrates. Structural analysis of B. subtilis ferrochelatase has shown that Tyr13 is an indirect ligand of Fe(2+) and a direct ligand of a copper mesoporphyrin product. A structure-based comparison revealed that Tyr13 aligns with a Met residue in the S. cerevisiae and human ferrochelatases. Tyr13 was changed to Met in the B. subtilis enzyme by site-directed mutagenesis. Enzymatic measurements showed that the modified enzyme inserted Co(2+) at a higher rate than the wild-type B. subtilis ferrochelatase, but it had lost the ability to use Cu(2+) as a substrate. Thus, the B. subtilis Tyr13Met ferrochelatase showed the same metal specificity as that of the ferrochelatases from S. cerevisiae and human.

AB - Ferrochelatase catalyzes the insertion of Fe(2+) into protoporphyrin IX. The enzymatic product heme (protoheme IX) is a well-known cofactor in a wide range of proteins. The insertion of metal ions other than Fe(2+) occurs rarely in vivo, but all ferrochelatases that have been studied can insert Zn(2+) at a good rate in vitro. Co(2+), but not Cu(2+), is known to be a good substrate of the mammalian and Saccharomyces cerevisiae ferrochelatases. In contrast, Cu(2+), but not Co(2+), has been found to be a good substrate of bacterial Bacillus subtilis ferrochelatase. It is not known how ferrochelatase discriminates between different metal ion substrates. Structural analysis of B. subtilis ferrochelatase has shown that Tyr13 is an indirect ligand of Fe(2+) and a direct ligand of a copper mesoporphyrin product. A structure-based comparison revealed that Tyr13 aligns with a Met residue in the S. cerevisiae and human ferrochelatases. Tyr13 was changed to Met in the B. subtilis enzyme by site-directed mutagenesis. Enzymatic measurements showed that the modified enzyme inserted Co(2+) at a higher rate than the wild-type B. subtilis ferrochelatase, but it had lost the ability to use Cu(2+) as a substrate. Thus, the B. subtilis Tyr13Met ferrochelatase showed the same metal specificity as that of the ferrochelatases from S. cerevisiae and human.

KW - Site-Directed

KW - Mutagenesis

KW - Ferrochelatase/chemistry/genetics/metabolism

KW - Copper/metabolism

KW - Cobalt/metabolism

KW - Bacillus subtilis/enzymology

KW - Bacterial Proteins/chemistry/genetics/metabolism

KW - Structure-Activity Relationship

KW - Substrate Specificity

KW - Tyrosine/chemistry/genetics/metabolism

U2 - 10.1007/s00775-010-0720-4

DO - 10.1007/s00775-010-0720-4

M3 - Article

VL - 16

SP - 235

EP - 242

JO - Journal of Biological Inorganic Chemistry

JF - Journal of Biological Inorganic Chemistry

SN - 1432-1327

IS - 2

ER -