Methemoglobin reductase pathway

This pathway mainly consists of:

• Cytochrome b5 (heme containing protein)
• Cytochrome b5 reductase (NADH-dependent methemoglobin reductase)
• NADH (Electron donor)
• NADPH
• NADPH-flavin reductase (NADPH-dependent methemoglobin reductase)
Two mechanism are involved in this pathway as there are two methemoglobin reductases in
erythrocytes; one is a NADH-dependent methemoglobin reductase which is also identified as
NADH-cytochrome b5 reductase and the other is NADPH-dependent methemoglobin reductase,
which is also known as NADPH-flavin reductase.
→ Methemoglobin reduction is majorly catalyzed by the NADH dependent cytochrome b5
methemoglobin reductase system.
→ The first component of the system is a flavoprotein named cytochrome b5 reductase. It is
also known as methemoglobin reductase.
→ The electrons used to reduce methemoglobin in this mechanism comes from glycolysis/
Embden meyerholf pathway, where glyceraldehyde-3-phosphate dehydrogenase reduces
NAD+ to NADH. This NADH acts as an electron donor.
→ Cytochrome b5 reductase transfers electrons from NADH to the second component of the
pathway, cytochrome b5.
→ Cyt b5oxidised + NADH → Cyt b5reduced + NAD+
→ Reduced cytochrome b5 then transfers the electrons to methemoglobin, reducing
Fe3+ back to the Fe2+state, thereby restoring hemoglobin to a fully functional state:
→ Hb - Fe3+ + Cyt b5reduced → Hb - Fe2+ + Cyt b5oxidised

→ The second mechanism in the methemoglobin reductase

pathway involves NADPH, which is produced by the
hexose monophosphate pathway.
→ In the hexose monophosphate pathway, the G6PD enzyme
converts glucose -6-phosphate to 6-phosphogluconate and
simultaneously reducing NADP+ to NADPH.
→ This NADPH uses the NADPH-dependent methemoglobin
reductase (NADPH-flavin reductase) to reduce the iron in
ferric state to ferrous state.
Significance of pathway

Erythrocytes are continuously subjected to oxidative stress and nitrite exposure, which results in
the spontaneous formation of metHb as hemoglobin is converted from its ferrous state to
methemoglobin in the ferric state. Methemoglobin can be fatal to the body. It reduces the oxygen
carrying capacity of the red blood cells as iron in its ferric state is unable to bind to oxygen, and
it also decreases the ability of the erythrocytes to release oxygen to the tissues. The efficiency of
the methemoglobin reductase pathway is that it ensures only trace quantities of methemoglobin
are normally present in erythrocytes. Hence, this pathway ensures that methemoglobin
accumulation in red cells is avoided and its concentration is maintained at less than 1% of the
total hemoglobin.

Effects of impairment of the pathway

Defects in this pathway can lead to methemoglobinemia. Methemoglobinemia occurs when

hemoglobin is oxidized to form methemoglobin, making it incapable of oxygen transport; if
severe, it leads to bluish discoloration of the skin and mucous membranes (cyanosis). There are
different types of methemoglobinemia. A deficiency in this methemoglobin reductase enzyme
can result in hereditary methemoglobinemia. Methemoglobin reductase is an enzyme used in the
methemoglobin pathway. which transfers electrons from NADH to methemoglobin, and usually
removes 95–99% of the methemoglobin produced. In this disease, particularly the type one of
this disease (also known as methemoglobin reductase deficiency), leads to the rate at which
methemoglobin converted to hemoglobin is unable to keep pace with the rate at which it is
formed. Hence, the increased amount of methemoglobin decreases the oxygen carrying capacity
of red blood cells, leading to severe hypoxia. Acquired methemoglobinemia can result from
being exposed to drugs and toxins, constant exposure to such toxins and oxidative stress lead to
overwhelming of the enzymes in the methemoglobin reductase pathway and hence, their
efficiency decreases and the oxygen carrying and releasing capacity of the red blood cells
decrease.