Hemerythin: structure, function, and spectroscopy

RC presentation

By Ansh Anant 19/04/2024

Hemerythrin: A Fascinating Oxygen Carrier Protein

• Hemerythrin is a metalloprotein found in marine invertebrates, particularly in

the coelomic uid of some worms and brachiopods. It plays a crucial role in
oxygen transport, similar to hemoglobin in vertebrates. This presentation will
delve into the structure, function, and spectroscopic properties of
hemerythrin, unveiling its unique mechanism for oxygen binding and release.
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 In invertebrate animals oxygen carrying proteins are often non-heme pigments,
hemerythrin and hemocyanin in place of the familiar hemoglobin.

• Both hemoglobin and hemerythin have iron as the oxygen carrying metal.

• Hemerythrin (as well as hemocyanin), it does not contain a heme group.

• Hemerythin combines with molecular oxygen in a ratio 2Fe: 02 rather than Fe:
O2 found in hemoglobin.

• In spite of these di erences both hemerythin and hemoglobin function

e ectively as oxygen carriers.

• Hemerythin is found in four di erent invertebrate phyla: sipunculids, poly-

chaetes, priapulids and bach
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 Structure of Hemerythrin

• Hemerythrin is a trimeric protein, meaning it consists of three identical

subunits.

• Each subunit folds into a β-propeller structure, forming a cavity that houses
the oxygen-binding site.

• The active site features two ferric iron (Fe(III)) ions bridged by a hydroxide
(OH-) group.

• Six amino acid residues, including histidine and glutamate, coordinate with
the iron ions, in uencing their electronic properties.
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Active site structure of Deoxyhemerythrin

Each monomeric unit contains an active site which has two high spin ferrous ions [Fe (Il)].

The ferrous ions are bridged together by a hydroxyl group and two carboxyl groups from
an aspartate residue and a glutamate residue of the protein chain.

One of the ferrous is hexacoordinated with an octahedral geometry and the other is
pentacoordinated with a distorted trigonal bipyramidal geometry.

The remaining coordination sites of hexacoordinated ferrous and pentacoordinated

ferrous are satis ed by three and two imidazole nitrogens respectively from
histidineresidues of the protein chain.

The hydroxyl group serves as a bridging ligand but also functions as a proton donor to
the 02 substrate.
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Active site structure of Oxyhaemerythrine

One monomeric unit of hemerythin binds one dioxygen.

The dioxygen adds only to the coordinatively unsaturated ferrous.

The dioxygen adds to hemerythrinin an oxidative manner resulting in the formation of two
Fe(Ill) centers and peroxide (022).

The oxidative addition is followed by the shifting of proton from the bridged OH to the
bound peroxide resulting in the formation of hydroperoxo (HO) group.

This proton-transfer resultin the formation of a single oxygen atom (u-oxo) bridge in
oxyhemerythrin.

The hydroperoxo group is hydrogen bonded with the u-oxo group

Binding of Dioxygen (02)
Function of Hemerythrin

• Hemerythrin functions as an oxygen transport protein in marine invertebrates.

• In the coelomic uid, hemerythrin binds to molecular oxygen (O2) in a
reversible manner.

• Oxygen binding involves the reduction of one Fe(III) ion to Fe(II) and the
formation of a peroxo (O2²⁻) bridge between the iron ions.

• Hemerythrin delivers oxygen to tissues by releasing it through the reverse

reaction, regenerating the Fe(III)-Fe(III) state.
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Spectroscopy of Hemerythrin
• Various spectroscopic techniques provide valuable insights into the structure
and function of hemerythrin.

• Electronic spectroscopy, particularly UV-visible (UV-Vis) absorption

spectroscopy, reveals characteristic peaks corresponding to electronic
transitions within the iron centers.

• Changes in the UV-Vis spectrum upon oxygen binding indicate alterations in

the electronic properties of the iron ions.

• Electron paramagnetic resonance (EPR) spectroscopy helps identify the

oxidation state and electronic con guration of the iron ions in both
deoxygenated and oxygenated hemerythrin.
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• The end products of the irreversible bimolecular oxidation of FeII species contain the
FeIlI—O—FeIII fragment. Given the facile formation of -oxodiiron(III) species, it is not
surprising that the Fe—O—Fe motif is incorporated into a variety of metalloproteins,
including the oxygen-carrier hemerythrin

• This -oxodiiron(III) moiety has a distinctive ngerprint that has made it easy to identify
this motif in proteins. Regardless of the number (4, 5, 6, or 7), geometry (tetrahedral,
square pyramidal, tetragonally distorted octahedral, or pentagonal bipyramidal), and
type of ligands (halide, RO-, RCOO-, aliphatic N, or aromatic N) around the iron center,
and of the Fe—O—Fe angle, the magnetic susceptibility at room temperature lies in
the range 1.5 to 2.0 Bohr magnetons per Felll—O—Fe lll group, equivalent to about
one unpaired electron. 81,82 In other words, the high-spin (S = ) iron centers are
strongly antiferromagnetically coupled. Other bridging groups, such as OH-, Cl-,
carboxylate, alkoxide, or phenoxide, give very weak coupling.83-86

• The asymmetric Fe—O stretch, vas(Fe—O), lies in the range 730 to 880 cm-1; in
multiply bridged complexes this mode is weak in the infrared region. The symmetric
vibration, vs(Fe—O), forbidden in the infrared region for linear, symmetric Fe—O—Fe
groups, occurs in the range 360 to 545 cm-1. The symmetric mode is usually,87a but
not always,87b observed by resonance Raman techniques upon irradiating on the low-
energy side of the Fe—O charge transfer band that occurs at about 350 nm.
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Conclusion
• Hemerythrin represents a fascinating example of how nature utilizes di erent
strategies for oxygen transport. Understanding its structure, function, and
spectroscopic properties contributes to our broader knowledge of
bioinorganic chemistry and the diverse adaptations of marine life. Further
research on hemerythrin may even lead to the development of novel
biomimetic materials for oxygen storage and delivery.

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