The C-terminal regions have an important role in the activity of the ferroxidase center and the stability of Chlorobium tepidum ferritin

Cristian Brito, Catalina Matias, Fernando D. González-Nilo, Richard K. Watt, Alejandro Yévenes

Resultado de la investigación: Article

3 Citas (Scopus)

Resumen

The recombinant Chlorobium tepidum ferritin (rCtFtn) is able to oxidize iron using ferroxidase activity but its ferroxidase activity is intermediate between the H-chain human ferritin and the L-chain human ferritin. The rCtFtn has an unusual C-terminal region composed of 12 histidine residues, as well as aspartate and glutamate residues. These residues act as potential metal ion ligands, and the rCtFtn homology model predicts that this region projects inside the protein cage. The rCtFtn also lacks a conserved Tyr residue in position 19. In order to know if those differences are responsible for the altered ferroxidase properties of rCtFtn, we introduced by site-directed mutagenesis a stop codon at position 166 and a Tyr residue replaced Ala19 in the gene of rCtFtn (rCtFtn 166). The rCtFtn166 keeps the canonical sequence considered important for the activity of this family of proteins. Therefore, we expected that rCtFtn 166 would possess similar properties to those described for this protein family. The rCtFtn 166 is able to bind, oxidize and store iron; and its activity is inhibit by Zn(II) as was described for other ferritins. However, the rCtFtn 166 possesses a decrease ferroxidase activity and protein stability compared with the wild type rCtFtn. The analysis of the Ala19Tyr rCtFtn shows that this change does not affect the kinetic of iron oxidation. Therefore, these results indicate that the C-terminal regions have an important role in the activity of the ferroxidase center and the stability of rCtFtn.

Idioma originalEnglish
Páginas (desde-hasta)211-220
Número de páginas10
PublicaciónProtein Journal
Volumen33
N.º3
DOI
EstadoPublished - 1 ene 2014

Huella dactilar

Chlorobium
Ceruloplasmin
Ferritins
Proteins
Iron
Mutagenesis
Metal ions
Genes
Ligands
Oxidation
Kinetics
Apoferritins

ASJC Scopus subject areas

  • Analytical Chemistry
  • Bioengineering
  • Biochemistry
  • Organic Chemistry

Citar esto

Brito, Cristian ; Matias, Catalina ; González-Nilo, Fernando D. ; Watt, Richard K. ; Yévenes, Alejandro. / The C-terminal regions have an important role in the activity of the ferroxidase center and the stability of Chlorobium tepidum ferritin. En: Protein Journal. 2014 ; Vol. 33, N.º 3. pp. 211-220.
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title = "The C-terminal regions have an important role in the activity of the ferroxidase center and the stability of Chlorobium tepidum ferritin",
abstract = "The recombinant Chlorobium tepidum ferritin (rCtFtn) is able to oxidize iron using ferroxidase activity but its ferroxidase activity is intermediate between the H-chain human ferritin and the L-chain human ferritin. The rCtFtn has an unusual C-terminal region composed of 12 histidine residues, as well as aspartate and glutamate residues. These residues act as potential metal ion ligands, and the rCtFtn homology model predicts that this region projects inside the protein cage. The rCtFtn also lacks a conserved Tyr residue in position 19. In order to know if those differences are responsible for the altered ferroxidase properties of rCtFtn, we introduced by site-directed mutagenesis a stop codon at position 166 and a Tyr residue replaced Ala19 in the gene of rCtFtn (rCtFtn 166). The rCtFtn166 keeps the canonical sequence considered important for the activity of this family of proteins. Therefore, we expected that rCtFtn 166 would possess similar properties to those described for this protein family. The rCtFtn 166 is able to bind, oxidize and store iron; and its activity is inhibit by Zn(II) as was described for other ferritins. However, the rCtFtn 166 possesses a decrease ferroxidase activity and protein stability compared with the wild type rCtFtn. The analysis of the Ala19Tyr rCtFtn shows that this change does not affect the kinetic of iron oxidation. Therefore, these results indicate that the C-terminal regions have an important role in the activity of the ferroxidase center and the stability of rCtFtn.",
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The C-terminal regions have an important role in the activity of the ferroxidase center and the stability of Chlorobium tepidum ferritin. / Brito, Cristian; Matias, Catalina; González-Nilo, Fernando D.; Watt, Richard K.; Yévenes, Alejandro.

En: Protein Journal, Vol. 33, N.º 3, 01.01.2014, p. 211-220.

Resultado de la investigación: Article

TY - JOUR

T1 - The C-terminal regions have an important role in the activity of the ferroxidase center and the stability of Chlorobium tepidum ferritin

AU - Brito, Cristian

AU - Matias, Catalina

AU - González-Nilo, Fernando D.

AU - Watt, Richard K.

AU - Yévenes, Alejandro

PY - 2014/1/1

Y1 - 2014/1/1

N2 - The recombinant Chlorobium tepidum ferritin (rCtFtn) is able to oxidize iron using ferroxidase activity but its ferroxidase activity is intermediate between the H-chain human ferritin and the L-chain human ferritin. The rCtFtn has an unusual C-terminal region composed of 12 histidine residues, as well as aspartate and glutamate residues. These residues act as potential metal ion ligands, and the rCtFtn homology model predicts that this region projects inside the protein cage. The rCtFtn also lacks a conserved Tyr residue in position 19. In order to know if those differences are responsible for the altered ferroxidase properties of rCtFtn, we introduced by site-directed mutagenesis a stop codon at position 166 and a Tyr residue replaced Ala19 in the gene of rCtFtn (rCtFtn 166). The rCtFtn166 keeps the canonical sequence considered important for the activity of this family of proteins. Therefore, we expected that rCtFtn 166 would possess similar properties to those described for this protein family. The rCtFtn 166 is able to bind, oxidize and store iron; and its activity is inhibit by Zn(II) as was described for other ferritins. However, the rCtFtn 166 possesses a decrease ferroxidase activity and protein stability compared with the wild type rCtFtn. The analysis of the Ala19Tyr rCtFtn shows that this change does not affect the kinetic of iron oxidation. Therefore, these results indicate that the C-terminal regions have an important role in the activity of the ferroxidase center and the stability of rCtFtn.

AB - The recombinant Chlorobium tepidum ferritin (rCtFtn) is able to oxidize iron using ferroxidase activity but its ferroxidase activity is intermediate between the H-chain human ferritin and the L-chain human ferritin. The rCtFtn has an unusual C-terminal region composed of 12 histidine residues, as well as aspartate and glutamate residues. These residues act as potential metal ion ligands, and the rCtFtn homology model predicts that this region projects inside the protein cage. The rCtFtn also lacks a conserved Tyr residue in position 19. In order to know if those differences are responsible for the altered ferroxidase properties of rCtFtn, we introduced by site-directed mutagenesis a stop codon at position 166 and a Tyr residue replaced Ala19 in the gene of rCtFtn (rCtFtn 166). The rCtFtn166 keeps the canonical sequence considered important for the activity of this family of proteins. Therefore, we expected that rCtFtn 166 would possess similar properties to those described for this protein family. The rCtFtn 166 is able to bind, oxidize and store iron; and its activity is inhibit by Zn(II) as was described for other ferritins. However, the rCtFtn 166 possesses a decrease ferroxidase activity and protein stability compared with the wild type rCtFtn. The analysis of the Ala19Tyr rCtFtn shows that this change does not affect the kinetic of iron oxidation. Therefore, these results indicate that the C-terminal regions have an important role in the activity of the ferroxidase center and the stability of rCtFtn.

KW - Bacterial ferritin

KW - Ferroxidase center

KW - Site-directed mutagenesis

KW - Transmission electron microscopy

UR - http://www.scopus.com/inward/record.url?scp=84902864885&partnerID=8YFLogxK

U2 - 10.1007/s10930-014-9552-3

DO - 10.1007/s10930-014-9552-3

M3 - Article

C2 - 24609571

AN - SCOPUS:84902864885

VL - 33

SP - 211

EP - 220

JO - Protein Journal

JF - Protein Journal

SN - 1572-3887

IS - 3

ER -