Aggregation of adult and fetal hemoglobin by ingested nitrate anions

Document Type: Research Paper


1 Laboratory of Microanalysis, Institute of Biochemistry and Biophysics (IBB), University of Tehran, P.O. Box 13145-1384, Tnarhe, Iran

2 Laboratory of Microanalysis, Institute of Biochemistry and Biophysics (IBB), University of Tehran, P.O. Box 13145-1384, Tnarhe, Iran; and Research Institute of Applied Science, Academic Center of Education, Culture and Research (ACECR), University of Shahid Beheshti, P.O. Box ,Tehran, Iran


The ingested nitrates sourced from tap water, food, chemicals and pharmaceuticals are
converted to nitrites in the body surfaces by bacteria and then, the nitrite ions can lead the
structural changing in hemoglobin. In the present work, aggregation of the purified hemoglobin
in adult (HbA) and in fetus or newborn (HbF) in the presence of nitrite ions were studied.
Hemoglobin aggregation was performed chemically in the presence of 10 mg/l nitrite ions and
examined by UV-Vis spectrophotometer at 360 nm wavelength. The extrinsic fluorimetric
measurements indicated that repulsive electrostatic interaction between nitrite anions and
negative charged groups of both types of HbA and HbF molecules leads to expose the
hydrophobic patch of the protein molecules. Moreover, the α-helix to β-strand transition in both
types of hemoglobins shown by circular dichroism support aggregation process among this
protein. However, at natural pH, the protonated amino group of Gly in HbF tends to bind to
nitrite anions more than the unprotonated forms of Val residue in HbA. The drastic slop of
aggregation plot and shorter lag time of HbF relative to HbA demonstrated more aggregation of
former protein.


Main Subjects

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