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Fetal hemoglobin is much less prone to DNA cleavage compared to the adult protein

Chakane, Sandeep LU ; Matos, Tiago LU ; Kettisen, Karin LU and Bulow, Leif LU (2017) In Redox Biology 12. p.114-120
Abstract

Hemoglobin (Hb) is well protected inside the red blood cells (RBCs). Upon hemolysis and when free in circulation, Hb can be involved in a range of radical generating reactions and may thereby attack several different biomolecules. In this study, we have examined the potential damaging effects of cell-free Hb on plasmid DNA (pDNA). Hb induced cleavage of supercoiled pDNA (sc pDNA) which was proportional to the concentration of Hb applied. Almost 70% of sc pDNA was converted to open circular or linear DNA using 10 µM of Hb in 12 h. Hb can be present in several different forms. The oxy (HbO2) and met forms are most reactive, while the carboxy-protein shows only low hydrolytic activity. Hemoglobin A (HbA) could easily induce... (More)

Hemoglobin (Hb) is well protected inside the red blood cells (RBCs). Upon hemolysis and when free in circulation, Hb can be involved in a range of radical generating reactions and may thereby attack several different biomolecules. In this study, we have examined the potential damaging effects of cell-free Hb on plasmid DNA (pDNA). Hb induced cleavage of supercoiled pDNA (sc pDNA) which was proportional to the concentration of Hb applied. Almost 70% of sc pDNA was converted to open circular or linear DNA using 10 µM of Hb in 12 h. Hb can be present in several different forms. The oxy (HbO2) and met forms are most reactive, while the carboxy-protein shows only low hydrolytic activity. Hemoglobin A (HbA) could easily induce complete pDNA cleavage while fetal hemoglobin (HbF) was three-fold less reactive. By inserting, a redox active cysteine residue on the surface of the alpha chain of HbF by site-directed mutagenesis, the DNA cleavage reaction was enhanced by 82%. Reactive oxygen species were not directly involved in the reaction since addition of superoxide dismutase and catalase did not prevent pDNA cleavage. The reactivity of Hb with pDNA can rather be associated with the formation of protein based radicals.

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organization
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type
Contribution to journal
publication status
published
subject
keywords
Adult hemoglobin, Cysteine, DNA cleavage, Fetal hemoglobin, Protein radicals, Supercoiled plasmid DNA
in
Redox Biology
volume
12
pages
7 pages
publisher
Elsevier
external identifiers
  • scopus:85013029047
  • wos:000403328700010
ISSN
2213-2317
DOI
10.1016/j.redox.2017.02.008
language
English
LU publication?
yes
id
c169981d-2878-42d2-b956-c3a349cbcbd8
date added to LUP
2017-03-01 07:39:36
date last changed
2018-01-07 11:53:20
@article{c169981d-2878-42d2-b956-c3a349cbcbd8,
  abstract     = {<p>Hemoglobin (Hb) is well protected inside the red blood cells (RBCs). Upon hemolysis and when free in circulation, Hb can be involved in a range of radical generating reactions and may thereby attack several different biomolecules. In this study, we have examined the potential damaging effects of cell-free Hb on plasmid DNA (pDNA). Hb induced cleavage of supercoiled pDNA (sc pDNA) which was proportional to the concentration of Hb applied. Almost 70% of sc pDNA was converted to open circular or linear DNA using 10 µM of Hb in 12 h. Hb can be present in several different forms. The oxy (HbO<sub>2</sub>) and met forms are most reactive, while the carboxy-protein shows only low hydrolytic activity. Hemoglobin A (HbA) could easily induce complete pDNA cleavage while fetal hemoglobin (HbF) was three-fold less reactive. By inserting, a redox active cysteine residue on the surface of the alpha chain of HbF by site-directed mutagenesis, the DNA cleavage reaction was enhanced by 82%. Reactive oxygen species were not directly involved in the reaction since addition of superoxide dismutase and catalase did not prevent pDNA cleavage. The reactivity of Hb with pDNA can rather be associated with the formation of protein based radicals.</p>},
  author       = {Chakane, Sandeep and Matos, Tiago and Kettisen, Karin and Bulow, Leif},
  issn         = {2213-2317},
  keyword      = {Adult hemoglobin,Cysteine,DNA cleavage,Fetal hemoglobin,Protein radicals,Supercoiled plasmid DNA},
  language     = {eng},
  month        = {08},
  pages        = {114--120},
  publisher    = {Elsevier},
  series       = {Redox Biology},
  title        = {Fetal hemoglobin is much less prone to DNA cleavage compared to the adult protein},
  url          = {http://dx.doi.org/10.1016/j.redox.2017.02.008},
  volume       = {12},
  year         = {2017},
}