Homo Sapiens Fetal Deoxyhemoglobin

Ubongabasi Asuquo '23 and Serena Liu '23


Contents:


I. Introduction




Homo sapiens fetal hemoglobin (HbF) is an oxygen carrier protein in the human fetus. It constitutes approximately 60 to 80 percent of total hemoglobin in the full-term newborn. It transports oxygen from the mother's bloodstream to the fetal circulation. From early pregnancy through the first six months of postnatal life, erythroid precursor cells produce HbF cells. HbF contains and as opposed to human adult hemoglobin (HbA), which consist of two alpha subunits and two beta subunits. Compared to HbA, HbF has a higher affinity of oxygen in vivo as a result of negatively charged phosphates, giving HbF the ability to bind to oxygen from maternal circulation. Here, we elaborate on the structure of human fetal deoxyhemoglobin. Deoxyhemoglobin is the form of hemoglobin that is not combined with oxygen. The four subunits are free, and because of that it is in the tense (T) state of hemoglobin. It is more soluble in this form, and it has a decreased binding affinity to 2,3-bisphosphoglycerate (2,3-DPG), an allosteric regulator, relative to deoxyHbA.


II. General Structure

HbF is an α2γ2 tetramer with a more compact overall structure. It can exist in both .

Alpha Subunits

The two α chains are termed αA and αF chains respectively. They are identical in amino acid composition, with both containing of alanine, leucine, and valine and none containing isoleucine. They also contain the same the tryptic peptide patterns and other subunit hydrizations. Out of fourteen possible tryptic peptides, twelve soluble peptides present in the αF chains showed similar amino acid compositions to equivalent peptides present in the αA chain.

Gamma Subunits

The γ subunits contain 146 residues each and are considered to be homologous to the β adult hemoglobin chains. Despite its close similarity, γ differs from β at 39 positions. The gamma subunit is responsible for the 70 fold enhanced strength of its tetramer-dimer dissociation constant. This is caused by the 18-amino-acid in the A helix, particularly . In addition, the γ subunit generally contains four isoleucine residues per polypeptide chain and its N-terminal sequence is often acetylated ending in . Unfortunately, the biological role of this acetylation is unknown at this time. In terms of protein charge, the positive charge of the gamma chain is relatively greater in the lys-104, ser-139, and ser-143. Some residues on side chains participate in hydrogen bonding; others are responsible for subunit contacts, internal and external surfaces and surface cervice. γ chains can also participate in disulfide bond formation as a result of on the chains. 

III. Adult vs. Fetal Hemoglobin

The three main functional differences between adult and fetal hemoglobin include: the increased oxygen affinity of HbF, the higher solubility of HbF in its deoxygenated form, and its higher tetramer-dimer dissociation constant. The higher solubility of deoxyHbF is a result of replacement of Glu by on the γ chains. This replacement lengthens the distance between the carboxyl group and the imidazole and thus weakens the electrostatic attraction between them. The primary sequence differences are mainly characterized in the , , , , , and the of the polypeptide chain (Table 1). These amino acid sequences hence contribute to the above mentioned functional differences.

Table 1. Sequence differences between beta chain and gamma chain (Frier and Perutz, 1977).
Table 1


IV. Subunit Assembly & Interactions

The ways through which the assembly of the α and γ subunits occur have been explored and are characterized to be slower when compared to the assembly of adult hemoglobin. This assembly is generally believed to occur by a multi-step mechanism, through which gamma chains assemble with alpha chains first with the help of : helix A, helix E, and helix H, and then stable γ2-chain dimers will be formed.

In addition, the dimers when formed are relatively stable unlike the dimers formed with beta chains, particularly α1γ1 versus α1β1. In addition, alpha chains exist in a monomer/dimer equilibrium and favor formation of monomers, whereas gamma chains are stablized in a monomer/tetramer equilibrium and favor the formation of tetramers. In general, when compared to beta adult hemoglobin chain, ala-51, thr-112, ile-116, glu-125, and asp-43 help at the to assemble the tetramer.

These interactions mentioned about are responsible for the increased stability of fetal hemoglobin. They are also responsible for the observed differences in the assembly of αγ versus αβ assembly in vitro and promotes the dissociation of gamma tetramers to dimers and monomers.

Interactions

on the gamma chain at the tetramer-dimer allosteric interaction contributes to the increased oxygen affinity of HbF by tightening the tetramer-dimer interface. When HbF binds oxygen, there is a rearrangement of subunits at the tetramer-dimer interface. This leads to the position of the R/T equilibrium changing, tightening the interface thus increasing the affinity for oxygen binding and gradually changing the protein configuration from a deoxy- to oxy- state. On the opposite end, this subtitution also lowers the response of HbF to the binding of 2,3-bisphosphoglycerate (2,3-DPG), which is an allosteric regulator synthesized in tissues with low ATP and high acid production.

In addition, two water molecules are also found at the α1γ1 interface of fetal deoxyhemoglobin. They form two , one between alpha thr-38 and gamma asp-99, and another between alpha thr-41 and gamma asp-99. The third water molecule which would usually be present in adult hemoglobin is not found in the α1γ1 contact of deoxyHbF. Instead, it is bonded to the carbonyl oxygen of Thr-41 and to N of Arg-40. 

V. Heme Group

Fetal hemoglobin contains with each chain containing one respectively. Each heme includes a Fe2+ bound in the center of a porphyrin, a large heterocyclic organic ring. The ferrous ion has two axial binding sites. The first site is occupied by an N of the closest histidine. The second site makes reversible binding to an oxygen atom possible through ion-induced dipole forces. The heme groups can bind and unbind oxygen, allowing oxygen to be transported (Figure 1).

Deoxy heme

Figure 1. Difference between oxy- and deoxy-heme. (A) Oxyhemoglobin has dioxygen (depicted as red) bound to the iron core of each heme group. The Fe2+ is centered in the porphyrin plane. (B) Deoxyhemoglobin is pictured (no oxygen is bound). The Fe2+ ion lies 0.4 Å outside of the porphyrin plane (Hemoglobin: Oxygen transport in mammals [Internet]. 2020).



VI. References

Adachi, K., Y. Zhao, T. Yamaguchi, and S. Surrey. 2000. Assembly of γ- with α-Globin Chains to Form Human Fetal Hemoglobin in Vitro and in Vivo. Journal of Biological Chemistry 275:1242412429. Elsevier.

Frier, J. A., and M. F. Perutz. 1977. Structure of human foetal deoxyhaemoglobin. J Mol Biol 112:97-112.

Hemoglobin: Oxygen transport in mammals [Internet]. 2020 [cited Dec 16 2021]. Available from: https://chem.libretexts.org/@go/page/188720

Kaufman, D. P., J. Khattar, and S. L. Lappin. 2021. Physiology, Fetal Hemoglobin. StatPearls. StatPearls Publishing, Treasure Island (FL).

Schroeder, W. A., J. R. Shelton, J. B. Shelton, and J. Cormick. 1962. Further sequences in the γ chain of human fetal hemoglobin. National Academy of Sciences .PNAS 48:284-287.

Schroeder, W. A., J. R. Shelton, J. B. Shelton, and J. Cormick. 1963. The amino acid sequence of the α chain of human fetal hemoglobin. Biochemistry 2:1353-1357.

Schroeder, W. A., J. R. Shelton, J. B. Shelton, J. Cormick, and R. T. Jones. 1963b. The Amino Acid Sequence of the γ Chain of Human Fetal Hemoglobin. Biochemistry 2:992-1008. American Chemical Society.

Back to Top