Myoglobin

Myoglobin is an iron- and oxygen-binding protein found in the muscle tissue of vertebrates in general and
in almost all mammals. It is related to hemoglobin, which is the iron- and oxygen-binding protein in
blood, specifically in the red blood cells. Myoglobin is only found in the bloodstream after muscle
injury. It is an abnormal finding, and can be diagnostically relevant when found in blood. [2]
Myoglobin is the primary oxygen-carrying pigment of muscle tissues.[3] High concentrations of
myoglobin in muscle cells allow organisms to hold their breath for a longer period of time. Diving
mammals such as whales and seals have muscles with particularly high abundance of myoglobin.[2]
Myoglobin is found in Type I muscle, Type II A and Type II B, but most texts consider myoglobin not to
be found in smooth muscle.
Myoglobin was the first protein to have its three-dimensional structure revealed by X-ray crystallography.
[4] This achievement was reported in 1958 by John Kendrew and associates.[5] For this discovery,
John Kendrew shared the 1962 Nobel Prize in chemistry with Max Perutz.[6] Despite being one of the
most studied proteins in biology, its physiological function is not yet conclusively established: mice
genetically engineered to lack myoglobin are viable, but showed a 30% reduction in volume of blood
being pumped by the heart during a contraction. They adapted to this deficiency through natural
reactions to inadequate oxygen supply (hypoxia) and a widening of blood vessels (vasodilation).[7] In
humans myoglobin is encoded by the MB gene.[8]
Meat color
Myoglobin contains hemes, pigments responsible for the color of red meat. The color that meat takes is
partly determined by the degree of oxidation of the myoglobin. In fresh meat the iron atom is the
ferrous state bound to a dioxygen molecule (O2). Meat cooked well done is brown because the iron
atom is now in the ferric (+3) oxidation state, having lost an electron. If meat has been exposed to
nitrites, it will remain pink because the iron atom is bound to NO, nitric oxide (true of, e.g., corned
beef or cured hams). Grilled meats can also take on a pink "smoke ring" that comes from the iron
binding to a molecule of carbon monoxide.[9] Raw meat packed in a carbon monoxide atmosphere
also shows this same pink "smoke ring" due to the same principles. Notably, the surface of this raw
meat also displays the pink color, which is usually associated in consumers' minds with fresh meat.
This artificially induced pink color can persist, reportedly up to one year.[10] Hormel and Cargill are
both reported to use this meat-packing process, and meat treated this way has been in the consumer
market since 2003.[11]
Role in disease
Myoglobin is released from damaged muscle tissue (rhabdomyolysis), which has very high concentrations
of myoglobin. The released myoglobin is filtered by the kidneys but is toxic to the renal tubular
epithelium and so may cause acute renal failure.[12] It is not the myoglobin itself that is toxic (it is a
protoxin) but the ferrihemate portion that is dissociated from myoglobin in acidic environments (e.g.,
acidic urine, lysosomes).
Myoglobin is a sensitive marker for muscle injury, making it a potential marker for heart attack in
patients with chest pain.[13] However, elevated myoglobin has low specificity for acute myocardial
infarction (AMI) and thus CK-MB, cTnT, ECG, and clinical signs should be taken into account to make
the diagnosis.
Structure and bonding
Myoglobin belongs to the globin superfamily of proteins, and as with other globins, consists of eight alpha
helices connected by loops. Human globin contains 154 amino acids.[15]

Myoglobin contains a porphyrin ring with an iron at its center. A proximal histidine group (His-94) is
attached directly to iron, and a distal histidine group (His-65) hovers near the opposite face.[15] The
distal imidazole is not bonded to the iron but is available to interact with the substrate O2. This
interaction encourages the binding of O2, but not carbon monoxide (CO), which still binds about 240
more strongly than O2.

The binding of O2 causes substantial structural change at the Fe center, which shrinks in radius and
moves into the center of N4 pocket. O2-binding induces "spin-pairing": the five-coordinate ferrous
deoxy form is high spin and the six coordinate oxy form is low spin and diamagnetic.

Synthetic analogues

Many models of myoglobin have been synthesized as part of a broad interest in transition metal dioxygen
complexes. A well known example is the picket fence porphyrin, which consists of a ferrous complex
of a sterically bulky derivative of tetraphenylporphyrin.[16] In the presence of an imidazole ligand,
this ferrous complex reversibly binds O2. The O2 substrate adopts a bent geometry, occupying the
sixth position of the iron center. A key property of this model is the slow formation of the -oxo dimer,
which is an inactive diferric state. In nature, such deactivation pathways are suppressed by protein
matrix that prevents close approach of the Fe-porphyrin assemblies.[17]
Structure
Myoglobin is a monomeric protein that has 153 amino acids residues. It consists of eight -helicies
connected through the turns with an Oxygen binding site. It has a globular structure. Myoglobin
contains a heme (prosthetic) group which is responsible for its main function (carrying of oxygen
molecules to muscle tissues). Myoglobin can exist in the oxygen free form, deoxymyoglobin, or in a
form in which the oxygen molecule is bound, called oxymyoglobin. Myoglobin is a protein found in
muscles that binds oxygen with its heme group like hemoglobin. Heme group consists of
protoporphyrin organic component and an iron atom located in its center. The heme group gives
muscle and blood their distinctive red color. The organic component consists of four pyrrole rings that
are linked by methine bridges. In addition, heme is responsible for the red color of the blood and
muscle. Oxidation of the iron atom (Fe2+ -> Fe3+) is mainly responsible for the color of muscle and
blood. At the center of protporphyrin, the iron atom is bonded to nitrogen atoms from four pyrrole
rings. The iron atom can form two additional bonds, one on each side of the heme plane. These
binding sites are called the fifth and sixth coordination sites. In myoglobin, the fifth coordination site is
occupied by the imidazole ring from a histidine residue on the protein. This hisitidine is referred to as
the proximal histidine. The sixth coordination site is available to bind oxygen. The iron atom in
deoxymyoglobin lies about four angstrom out of the plane of the protoporphyrin plane because it is
too big in that form to fit into the well defined hole.
The normal oxidation state of an iron atom has a positive two charge (ferrous ion) instead of three charge
(ferric ion) and it is too large to fit into the plane of protoporphyrin. Thus, an ferrous ion often 0.4A
away from porphyrin plane. However, when iron oxidized from ferrous ion (Fe2+) to ferric ion (Fe3+),
because the lost of one extra electron, forces between protons and electrons increases so that the
electron cloud will penetrate more towards to the nucleus. As a result, the ferric ion (Fe3+) has a
smaller size then ferrous ion (Fe2+) and fits into the protoporphyrin plane when it attaches to an
oxygen.
When oxygen leaves the myoglobin, it leaves as dioxygen rather than superoxide. This is because
superoxide can be damaging to many biological process, and in the leaving of superdioxide, the iron
ion will be in the ferric state which stops biding oxygen.

The distal histidine amino acid from the hemoglobin protein molecule further stabilizes the O2 molecule
by hydrogen-bonding interactions.
Myoglobin is a protein molecule that has a similar structure and function to hemoglobin. It is a smaller
monomer of polypeptide structure, a globular protein with amino acids and prosthetic heme group
binds to proximal histidine group while a distal histidine group interact on the other side of the plane.
It binds and stores oxygen without concerning cooperativity. Most importantly, it is the first protein
structure to be studied.

Myoglobin follows the Michaelis-Menten Kinetic graph. (as seen from the graph above) It follows the
Michaelis-Menten kinetics because it is a simple chemical equilibrium.
Function
The binding affinities for oxygen between myoglobin and hemoglobin are important factors for their
function. Both myoglobin and hemoglobin binds oxygen well when the concentration of oxygen is
really high (E.g. in Lung), however, hemoglobin is more likely to release oxygen in areas of low
concentration (E.g. in tissues). Since hemoglobin binds oxygen less tightly than myoglobin in muscle
tissues, it can effectively transport oxygen throughout the body and deliver it to the cells. Myoglobin,

on the other, would not be as efficient in transferring oxygen. It does not show the cooperative
binding of oxygen because it would take up oxygen and only release in extreme conditions. Myoglobin
has a strong affinity for oxygen that allows it to store oxygen in muscle effectively. This is important
when the body is starve for oxygen, such as during anaerobic exercise. During that time, carbon
dioxide level in blood streams is extremely high and lactic acid concentration build up in muscles.
Both of these factors cause myoglobin (and hemoglobins) to release oxygen, for protecting the body
tissues from getting damaged under harsh conditions. If the concentration of myoglobin is high within
the muscle cells, the organism is able to hold the breath for a much longer period of time.
Myoglobin, an iron-containing protein in muscle, receives oxygen from the red blood cells and transports
it to the mitochondria of muscle cells, where the oxygen is used in cellular respiration to produce
energy. Each myoglobin molecule has one heme prosthetic group located in the hydrophobic cleft in
the protein. The function of myoglobin is notable from Millikan's review (1) in which he put together
an accomplished study to establish that myoglobin is formed adaptively in tissues in response to
oxygen needs and that myoglobin contributes to the oxygen supply of these tissues. Oxymyoglobin
regulates both oxygen supply and utilization by acting as a scavenger of the bioactive molecule nitric
oxide. Nitric oxide is generated continuously in the myocyte. Oxymyoglobin reacts with NO to form
harmless nitrates, with concomitant formation of ferric myoglobin, which is recycled through the
action of the intracellular enzyme metmyoglobin reductase. Flogel (2) conducted a study that showed
how the interaction of NO and oxymyoglobin controls cardiac oxygen utilization. Function[edit]
The binding affinities for oxygen between myoglobin and hemoglobin are important factors for their
function. Both myoglobin and hemoglobin binds oxygen well when the concentration of oxygen is
really high (E.g. in Lung), however, hemoglobin is more likely to release oxygen in areas of low
concentration (E.g. in tissues). Since hemoglobin binds oxygen less tightly than myoglobin in muscle
tissues, it can effectively transport oxygen throughout the body and deliver it to the cells. Myoglobin,
on the other, would not be as efficient in transferring oxygen. It does not show the cooperative
binding of oxygen because it would take up oxygen and only release in extreme conditions. Myoglobin
has a strong affinity for oxygen that allows it to store oxygen in muscle effectively. This is important
when the body is starve for oxygen, such as during anaerobic exercise. During that time, carbon
dioxide level in blood streams is extremely high and lactic acid concentration build up in muscles.
Both of these factors cause myoglobin (and hemoglobins) to release oxygen, for protecting the body
tissues from getting damaged under harsh conditions. If the concentration of myoglobin is high within
the muscle cells, the organism is able to hold the breath for a much longer period of time.
Myoglobin, an iron-containing protein in muscle, receives oxygen from the red blood cells and transports
it to the mitochondria of muscle cells, where the oxygen is used in cellular respiration to produce
energy. Each myoglobin molecule has one heme prosthetic group located in the hydrophobic cleft in
the protein. The function of myoglobin is notable from Millikan's review (1) in which he put together
an accomplished study to establish that myoglobin is formed adaptively in tissues in response to
oxygen needs and that myoglobin contributes to the oxygen supply of these tissues. Oxymyoglobin
regulates both oxygen supply and utilization by acting as a scavenger of the bioactive molecule nitric
oxide. Nitric oxide is generated continuously in the myocyte. Oxymyoglobin reacts with NO to form
harmless nitrates, with concomitant formation of ferric myoglobin, which is recycled through the
action of the intracellular enzyme metmyoglobin reductase. Flogel (2) conducted a study that showed
how the interaction of NO and oxymyoglobin controls cardiac oxygen utilization.

Hemerythrin
Hemerythrin (also spelled haemerythrin; from Greek words = blood and = red) is an
oligomeric protein responsible for oxygen (O2) transport in the marine invertebrate phyla of
sipunculids, priapulids, brachiopods, and in a single annelid worm genus, Magelona. Myohemerythrin
is a monomeric O2-binding protein found in the muscles of marine invertebrates. Hemerythrin and
myohemerythrin are essentially colorless when deoxygenated, but turn a violet-pink in the
oxygenated state.
Hemerythrin does not, as the name might suggest, contain a heme. The names of the blood oxygen
transporters hemoglobin, hemocyanin, hemerythrin, do not refer to the heme group (only found in
globins), instead these names are derived from the Greek word for blood.
O2 binding mechanism[edit]

The mechanism of dioxygen binding is unusual. Most O2 carriers operate via formation of dioxygen
complexes, but hemerythrin holds the O2 as a hydroperoxide. The site that binds O2 consists of a pair
of iron centres. The iron atoms are bound to the protein through the carboxylate side chains of a

glutamate and aspartates as well as through five histidine residues. Hemerythrin and
myohemerythrin are often described according to oxidation and ligation states of the iron centre:
Fe2+OHFe2+

deoxy (reduced)

Fe2+OHFe3+

semi-met

Fe3+OFe3+OOH-

oxy (oxidized)

Fe3+OHFe3+ (any other ligand)

met (oxidized)

The uptake of O2 by hemerythrin is accompanied by two-electron oxidation of the diferrous centre to

produce a hydroperoxide (OOH-) complex.

Hemocyanin
Hemocyanins (also spelled haemocyanins) are proteins that transport oxygen throughout the bodies of
some invertebrate animals. These metalloproteins contain two copper atoms that reversibly bind a
single oxygen molecule (O2). They are second only to hemoglobin in frequency of use as an oxygen
transport molecule. Unlike the hemoglobin in red blood cells found in vertebrates, hemocyanins are
not bound to blood cells but are instead suspended directly in the hemolymph. Oxygenation causes a
color change between the colorless Cu(I) deoxygenated form and the blue Cu(II) oxygenated form.
Structure and mechanism
The underside of the carapace of a Cancer productus crab. The purple coloring is caused by
hemocyanin.
Although the respiratory function of hemocyanin is similar to that of hemoglobin, there are a significant
number of differences in its molecular structure and mechanism. Whereas hemoglobin carries its iron
atoms in porphyrin rings (heme groups), the copper atoms of hemocyanin are bound as prosthetic
groups coordinated by histidine residues. It has been noted that species using hemocyanin for oxygen
transportation include crustaceans living in cold environments with low oxygen pressure. Under these
circumstances hemoglobin oxygen transportation is less efficient than hemocyanin oxygen
transportation.[6] Nevertheless there are also terrestrial arthropods using hemocyanin, notably
spiders and scorpions, that live in warm climates.
Most hemocyanins bind with oxygen non-cooperatively and are roughly one-fourth as efficient as
hemoglobin at transporting oxygen per amount of blood. Hemoglobin binds oxygen cooperatively due
to steric conformation changes in the protein complex, which increases hemoglobin's affinity for
oxygen when partially oxygenated. In some hemocyanins of horseshoe crabs and some other species
of arthropods, cooperative binding is observed, with Hill coefficients of 1.6 - 3.0. Hill coefficients vary
depending on species and laboratory measurement settings. Hemoglobin, for comparison, has a Hill
coefficient of usually 2.8 - 3.0. In these cases of cooperative binding hemocyanin was arranged in
protein sub-complexes of 6 subunits (hexamer) each with one oxygen binding site; binding of oxygen
on one unit in the complex would increase the affinity of the neighboring units. Each hexamer
complex was arranged together to form a larger complex of dozens of hexamers. In one study,
cooperative binding was found to be dependent on hexamers being arranged together in the larger
complex, suggesting cooperative binding between hexamers. Hemocyanin oxygen-binding profile is
also affected by dissolved salt ion levels and pH.[7]
Hemocyanin is made of many individual subunit proteins, each of which contains two copper atoms and
can bind one oxygen molecule (O2). Each subunit weighs about 75 kilodaltons (kDa). Subunits may be
arranged in dimers or hexamers depending on species; the dimer or hexamer complex is likewise
arranged in chains or clusters with weights exceeding 1500 kDa. The subunits are usually
homogeneous, or heterogeneous with two variant subunit types. Because of the large size of
hemocyanin, it is usually found free-floating in the blood, unlike hemoglobin.[8]
Hexamers are characteristic of arthropod hemocyanins.[9] A hemocyanin of the tarantula Eurypelma
californicum[2] is made up of 4 hexamers or 24 pepide chains. A hemocyanin from the house
centipede Scutigera coleoptrata[10] is made up of 6 hexamers or 36 chains. Horseshoe crabs have an
8-hexamer (i. e. 48-chain) hemocyanin. Simple hexamers are found in the spiny lobster Panulirus
interruptus and the isopod Bathynomus giganteus.[11] Peptide chains in crustaceans are about 660

amino acid residues long, and in chelicerates they are about 625. In the large complexes there is a
variety of variant chains, all about the same length; pure components do not usually self-assemble.
Catalytic activity
Hemocyanin is homologous to the phenol oxidases (e.g. tyrosinase) since both enzymes sharing type 3
Cu active site coordination. Hemocyanin also exhibits phenol oxidase activity, but with slowed kinetics
from greater steric bulk at the active site. Partial denaturation actually improves hemocyanins phenol
oxidase activity by providing greater access to the active site.[12]
Spectral properties
Spectroscopy of oxyhemocyanin shows several salient features:
resonance Raman spectroscopy shows symmetric binding
UV-Vis spectroscopy shows strong absorbances at 350 and 580 nm.
OxyHc is EPR-silent indicating the absence of unpaired electrons
Infrared spectroscopy shows (O-O) of 755 cm-1

(1) rules out a mononuclear peroxo complex (2) does not match with the UV-Vis spectra of mononuclear
peroxo and Kenneth Karlin's trans-peroxo models.[13] (4) shows a considerably weaker O-O bond
compared with Karlin's trans-peroxo model.[13]
On the other hand, Nobumasa Kitajima's model shows (O-O) of 741 cm-1 and UV-Vis absorbances at
349 and 551 nm, which agree with the experimental observations for oxyHc.[

Iron-sulfur protein
Iron-sulfur proteins are proteins characterized by the presence of iron-sulfur clusters containing sulfidelinked di-, tri-, and tetrairon centers in variable oxidation states. Iron-sulfur clusters are found in a
variety of metalloproteins, such as the ferredoxins, as well as NADH dehydrogenase, hydrogenases,
Coenzyme Q - cytochrome c reductase, Succinate - coenzyme Q reductase and nitrogenase.[1] Ironsulfur clusters are best known for their role in the oxidation-reduction reactions of mitochondrial
electron transport. Both Complex I and Complex II of oxidative phosphorylation have multiple Fe-S
clusters. They have many other functions including catalysis as illustrated by aconitase, generation of
radicals as illustrated by SAM-dependent enzymes, and as sulfur donors in the biosynthesis of lipoic
acid and biotin. Additionally some Fe-S proteins regulate gene expression. Fe-S proteins are
vulnerable to attack by biogenic nitric oxide.

The prevalence of these proteins on the metabolic pathways of most organisms leads some scientists to
theorize that iron-sulfur compounds had a significant role in the origin of life in the Iron-sulfur world
theory.
Structural motifs[edit]

In almost all Fe-S proteins, the Fe centers are tetrahedral and the terminal ligands are thiolato sulfur
centers from cysteinyl residues. The sulfide groups are either two- or three-coordinated. Three distinct
kinds of Fe-S clusters with these features are most common.
2Fe-2S clusters
The simplest polymetallic system, the [Fe2S2] cluster, is constituted by two iron ions bridged by two
sulfide ions and coordinated by four cysteinyl ligands (in Fe2S2 ferredoxins) or by two cysteines and
two histidines (in Rieske proteins). The oxidized proteins contain two Fe3+ ions, whereas the reduced
proteins contain one Fe3+ and one Fe2+ ion. These species exist in two oxidation states, (FeIII)2 and
FeIIIFeII.
4Fe-4S clusters
A common motif features a four iron ions and four sulfide ions placed at the vertices of a cubane-type
structure. The Fe centers are typically further coordinated by cysteinyl ligands. The [Fe4S4] electrontransfer proteins ([Fe4S4] ferredoxins) may be further subdivided into low-potential (bacterial-type)

and high-potential (HiPIP) ferredoxins. Low- and high-potential ferredoxins are related by the following
redox scheme:
In HiPIP, the cluster shuttles between [2Fe3+, 2Fe2+] (Fe4S42+) and [3Fe3+, Fe2+] (Fe4S43+). The
potentials for this redox couple range from 0.4 to 0.1 V. In the bacterial ferredoxins, the pair of
oxidation states are [Fe3+, 3Fe2+] (Fe4S4+) and [2Fe3+, 2Fe2+] (Fe4S42+). The potentials for this
redox couple range from -0.3 to -0.7 V. The two families of 4Fe-4S clusters share the Fe4S42+
oxidation state. The difference in the redox couples is attributed to the degree of hydrogen bonding,
which strongly modifies the basicity of the cysteinyl thiolate ligands. A further redox couple, which is
still more reducing than the bacterial ferredoxins is implicated in the nitrogenase.
Some 4Fe-4S clusters bind substrates and are thus classified as enzyme cofactors. In aconitase, the Fe-S
cluster binds aconitate at the one Fe centre that lacks a thiolate ligand. The cluster does not undergo
redox, but serves as a Lewis acid catalyst to convert aconitate to isocitrate. In radical SAM enzymes,
the cluster binds and reduces S-adenosylmethionine to generate a radical, which is involved in many
biosyntheses.[2]
3Fe-4S clusters
Proteins are also known to contain [Fe3S4] centres, which feature one iron less than the more common
[Fe4S4] cores. Three sulfide ions bridge two iron ions each, while the fourth sulfide bridges three iron
ions. Their formal oxidation states may vary from [Fe3S4]+ (all-Fe3+ form) to [Fe3S4]2- (all-Fe2+
form). In a number of iron-sulfur proteins, the [Fe4S4] cluster can be reversibly converted by oxidation
and loss of one iron ion to a [Fe3S4] cluster. E.g., the inactive form of aconitase possesses an [Fe3S4]
and is activated by addition of Fe2+ and reductant.
Other Fe-S clusters
More complex polymetallic systems are common. Examples include both the 8Fe and the 7Fe clusters in
nitrogenase. Carbon monooxide dehydrogenase and the [FeFe]-hydrogenase also feature unusual Fe-S
clusters. A special 6 cysteine-coordinated [Fe4S3] cluster was found in oxygen-tolerant membranebound [NiFe] hydrogenases.[3][4]