Bioinorganic Chemistry

Porphyrins and Related Complexes

in Bioinorganic Molecules
• A porphine ring has a square planar geometry with a
“pocket” in the center.
• A metalloporphyrin complex can result by
incorporating a metal atom into the pocket .
– Axial sites are available for other ligands.
• Structure, specificity, and reactivity are changed by
differing the side chains, metal ions, and surrounding
species.
Porphyrin ring is
rigid due to long π
delocalisation and
its cavity size is
good for first row
transition metal
In living systems iron has three well characterised systems:
1. Protiens that contain one or mor eporphyrin ring such as
haemoglobin, myoglobin and cytochrome P450.
2. Protiens that contain non-heme iorn such as
rubredoxin,ferredoxins, nitrogenase etc.
3. The non heme diiron oxo briged compounds such as
hemerythrine, ribonucleotide reductase, methane monooxygenase.
 Hemoglobin and Myoglobin
• Haemoglobin is made up of four globin protein subunits (α and β).
– Each protein partially encloses a heme group.
A myglobin is euivalent to one single unit of haemoglobin or a haemoglobin is
euivalent to four myglobin units.
• Each heme group is in a porphyrin pocket.
– One axial position of the iron is bound to an imidazole nitrogen from the
protein.
– One axial position is available/vacant.
• Dissolved O2 can bind reversibly to this axial position.
• In hemoglobin, the Fe(II) does not become oxidized to Fe(III) or Fe(IV).
• A free heme also oxidizes in an aqueous environment.
– Why doesn’t oxidation occur in hemoglobin by O2 or H2O?

The function of hemoglobin is to bind O2 at high oxygen pressure and carry it

through the blood to needed areas (and myoglobin for storage).
Hb + 4O2 → Hb(O2)4
Hb(O2)4 + 4Mb → 4Mb(O2) + Hb
 Structure of hemoglobin
• Hb is a spherical molecule consisting of 4 peptide subunits (globins) =
quartenary structure
• Hb of adults is a tetramer consisting of 2 α-(141 amino acids) and 2 β-(146
amino acids ) globins → each globin contains 1 heme group with a
central Fe2+ ion (ferrous ion)
• Oxygen transfer and storage agents in the blood and muscle tissue.
– Hemoglobin transports oxygen (O2) from the lungs/gills to tissues
and muscles.
– Myoglobin stores oxygen (O2) in the muscles and tissues.
Oxygen commonly transfers from the hemoglobin to the myoglobin
for later use.
 Due to coupplig of one unpaired electron on
Fe(III) and one from superoxide.

In deoxy haemoglobin Fe(II) is high spin so the unpaired electron in dx2-y2 repulses the
lone pair of electron on nitrogen atom and effectively radious of Fe(II) increases so it
doesnot fit into the porphyrin ring core and comes upward of the ring by 40 pm in the
direction of hystidine ring and porphyrin ring takes a domed shape. The Fe(II) centre is
square pyramidal in deoxy form.
Although O2 is not a strong ligand the
coordination of it trans to histidine as sixth
ligans alters the ligand strength and now it
becomes a high spin centre. So now dx2-y2 is
empty, so no repulsion with nitrogen lone
pair and also as it gets oxidised to Fe(III)
radious reduces by 17 pm so it fits into the
porphyrin core, so moves to the core, as a
result the coordinated histidine also comes
closer to the ring. Fe(III) has octahedral
geometry.O2 binds in bent fashion. Fe-O-O
angle is 150o. There is a strong evidence of
strong H- bonding of the bonded dioxygen
with a distal hystidine moiety. It’s a
superoxide now to be precise.
The protein structure of the 4 subunit of haemoglobin consists of peptide backbone
with several side chains ( these side chain contains non polar, cationic :-NH3+ and
anionic: -COO- groups). Due to presence of these charged side chains there occurs
electrostatic interaction and forms salt bridge and thus 4 subunits are held
together. These salt bridges are believed to provide some starin into it, that’s why
is it called T tense state. The movement of the Fe atom and histidine ring breaks
some of the salt bridge and thus some strain is released in oxy-Haemoglobin; thus
it’s called Relaxed or R-state.

After binding one O2 molecule as the strain is slightly released, it helps to bind
another O2 molecule to another Fe(II) centre in another subunit, as then some
more strain will be released. In this was all for deoxy-haemoglobin units gets
transformed into oxy-haemoglobin. This phenomenon is called “ Cooperative
Effect”.
Hb + 4O2 → Hb(O2)4
K1<K2<K3<K4
Hb(O2)4 + 4Mb → 4Mb(O2) + Hb
Conversely as one O2 molecule is removed from oxyhaemoglobin the reverse
conformational change occurs and successive decrease its affinity for oxygen.
Therefore initial removal of O2 molecule from one subunit triggers the removal of
other O2 molecules. This is also called cooperative effect.
K1<K2<K3<K4
Naked heme

The hydrocarbon environment round the iron

has a low dielectric constant and is hydrophobic,
therefore acts as a non-polar environment and
hematin provides non-aq. environment. It also provides
streric hindrance and doesn’t allow hematin
formation. Hematin is unable to transport oxygen.
Model system : picket fence porphyrin
 Oxygen binding curve
Hymoglobin has relatively higher affinity for dioxygen at higher
partial pressure and myoglobin has it in lower partial pressure.
Hb + 4O2 → Hb(O2)4 @ lungs
Hb(O2)4 + 4Mb → 4Mb(O2) + Hb @ muscle tissues

At low partial pressures of O2, Mb

has a much greater affinity for O2.

[Mb(O 2 )]
K Mb =
[Mb][O 2 ]

[ Hb(O2 ) 4 ]
K Hb =
[ Hb][O2 ]2.8

2.8 : called Hill’s constant.

Increased acidity favors the release of O2 from Hb(O2)4
In muscle tissue CO2 is present; acidic in nature.

CO2 is transported to lungs as soluble HCO3-, produced by enzyme carbonic anhydrase

as H2CO3 ; which dissociated to HCO3- and H+. This dissociation is favoured by deoxy-
Haemoglobin which acts as a buffer by picking up the protons. HCO3- travels to lungs
in blood serum and there release of proton from haemoglobic acid forms H2CO3 from it,
which gets converted to water and CO2 by same enzyme carbonic anhydrase and CO2
gets exhaled out and regerated haemoglobin again binds with di oxygen.

Thus in tissue cell with increasing partial pressure of CO2 i.e. which increasing CO2
concentration acidity increases i.e. pH increases i.e. protonation of haemoglobin
increases so Dioxygen gets out of oxy-Haemoglobin and consequently partial
saturation with oxygen decreases.
 Poisoning effect of CO &CN-

O2 is not a strong ligand. It is a π acceptor ligand.

CO, CN-, NO, PF3 these are strong ligands and even stronge π acceptor ligands .

CO binds (200 times stronger) competitively at the same site of Haemoglobin as does
O2 and also shows similar Co-operative and Bohr effect.The amount of O2 that can be
carried out by the remaining Fe2+ centres of the haemoglobin molecule is reduced if CO
is bound one of the subunits. Thus O2 transfer becomes tougher. It prevents the transport
of O2 and causes death due to Asphyxia (body deprived of oxygen; suffocation).
 Hemerythrin
1. Non-heme Fe containing oxygen career protein: Eight subunits: 113 amino acids
2. Major difference with haemoglobin: no co-oerativity between subunits, one dioxygen molecule
binds between two Fe(II) centre.
3. Fe(II) centres in deoxyhemerythrine are high spin and strongly antiferromagnetically coupled
through Fe(II)-O-Fe(II) bridge.
4. Dioxygen binds to coordinatively unsaturated Fe(II) centre.
5. During dioxygen binding Fe(II) centres looses one electron each and becomes low spin Fe(III);
those 2 electrons are taken up by dioxygen and gets converted to peroxide (O2)2- . The proton of
shifts to bound peroxide foring HOO- group.
6. Two Fe(III) centres are differrent. Presence of peroxide proved by raman spectroscopy O-O
stretching frequecy @ 845 cm-1.Two centres are antiferromagnetically coupled; overall
diamagnetic; EPR inactive.
7. When oxidised called met-hemeerythrine, contains oxo- not hydroxo-, cant bind oxygen.
 Hemocyanin
1. Non-heme Cu containg oxygen career protein.
2. Deoxy form colourless, Cu(I) centres are 460 pm apart. Triagonal planner geometry.
3. Has several sub units; shows co-operativity.
4. Oxy from shows blue colour due to LMCT [O2- to Cu2+]. Cu(II) centres comes closer; 360 pm
apart. Square pyramidal geometry.
5. During formation of oxy form each Cu(I) centre loses one electron and forms Cu(II) centre and
the 2 electrons are taken by dioxygen and forms peroxide.
6. Presence of peroxide proved by Raman spectroscopy.
 Cytochromes (a, b &c)
Cytochromes are hemoproteins containing heme groups and are primarily responsible for
the generation of ATP via electron transport. Serves as electron carrier. In Cytochrome a, b
&c all sites are filled up so neither oxygen nor poisonous CO can get attached to it, so inert
to these thing.
 Cytochrome-c oxidase

CuA

Heme a a3-CuB
CuB
 Cytochrome c oxidase which binds the inner
membrane of mitochondreon contains
cytochrome-a, cytochrome-a3 and two Cu
centres CuA (with oxidation state +2) and CuB
(with oxidation state +1), electron gets
transferres from cytochrome-c to CuA & then to
cytochrome-a. Cytochrome-a3 and CuB provide
sites for dioxygen binding and conversion of O2
to H2O. Cytochrome-a3 and CuB are five and
three coordinated respectively and therefore
binds dioxygen. The over all reaction is as
described. This also indicates cytochrome chain
involves transport of proton along with electron
along mitochondrion membrane. Cytochrome –
a3 centre is five membered so its oxidised form
can bind to CN- and thus occurs cyanide
poisoning, which leads to stopping electron
transfer.
Organisation of redox active
centre of cytochrome c oxidase

4cyt c2+ + O2+ 8H+inside 4cyt c3+ +2H2O +4H+outside

Cytochrome P450: hydroxylation (monooxygenase)
Catalytic Cycle of Cytochrome P450

Active site of cytochrome P450 is

different from haemoglobin or
myoglobin by the following:
1.Fe is present in +3 oxidation
state and is low spin.
2.octahedral, one S atom of
cysteine is coordinated instead of
histidine.
3. Sixth site is occupied by water.
Methane Monooxygenase: methane to methanol
 Iron-sulfur protein
•Non-heme protein
•Does electron transfer inplants and bacteria; acts as a reducing
agent( in biological redox reactions).
•Contains one, two, three, four or eight Fe atoms, each Fe atom is
coordinated to four S atom in approximately tetrahedral manner.
•Fe atoms are high spin.
•Involved in photosynthesis, nitrogen fixation etc.
Rubredoxin
•Simplest non-heme iron protein; only one Fe centre; transfers
one electron
•Distorted tetrahedral; coordinated to 4 S atoms of cystein amino
acid of globin chain.
•Fe is in +3 oxidation state.
•Non-labile S coordination; inorganic brigring S2- are labile,
liberated as H2S by treatment of mineral acids.
 +e

-e

Both Fe centre are in high spin state

Ferrodoxins
•Three major types: 2Fe-2S or Fe2S2, 3Fe-4S or Fe3S4 and Fe4-
S4 or Fe4S4.
 2Fe-2S or Fe2S2

S=0 S=1/2

•Called plant ferridoxins; both Fe(III) & Fe(II) centres are high spin in all forms. Both Fe centres
are brigged via two labile sulfide ions and also coordinated with two non labile cystein sulfer
atoms.
•One elec. Transfer protein; reduced from antiferromagnetically coupled ; diamagnetic; S=0; EPR
inactive; oxidised form antiferromagnetically coupled; S=1/2; epr active.
•mossbauer spectroscopy says both the Fe centres are same so overall charge on each Fe centre is
fractional and is +2.5 in reduced form.
RIESKE protein
 3Fe-4S or Fe3S4

S=5/2 S=2
•In oxidised form 3 Fe (III) & in reduced form 2 Fe(III) & 1 Fe(II) centres. Cubane
structure with one atom missing.
•Both Fe(III) & Fe(II) centres are high spin in all forms. Each Fe centre is brigged via
three labile sulfide ions and also coordinated with one non labile cyystein sulfer atom.
•One elec. Transfer protein;

in reduced form two Fe(III) centres are antiferromagnetically coupled ; third one is
free so S=5/2; EPR active;
in oxidised form two Fe(III) centres are antiferromagnetically coupled; Fe(II) high
spin centre is free, has 4 unpaired electron so S=2; epr inactive.
 4Fe-4S or Fe4S4
•Most common and most stable. Cubane structure.
•In oxidised form 2 Fe (III),2 Fe (II) & in reduced form 1 Fe(III) & 3 Fe(II) centres.
•Both Fe(III) & Fe(II) centres are high spin in all forms. Each Fe centre is brigged via three labile
sulfide ions and also coordinated with one non labile cyystein sulfer atom.
•One elec. Transfer protien;
in reduced form two Fe(III) centres are antiferromagnetically coupled ; also both Fe(II)
centres are also antiferromagnetically coupled. so S=0;diamagnetic; EPR inactive;
in oxidised form two Fe(II) centres are antiferromagnetically coupled; among the other two
Fe(III) high spin centre has 5 unpaired electrons and Fe(II) centre has 4 unpaired electron , they
are also anti ferromagnetically coupled with overall S=1/2; EPR active.
.Mossbauer spectroscopy says all Fe centres are similar; so overall charge on each Fe centre on
oxidised form is +2.5 and is +2.25 on reduced form.
 Nitrogen fixation

On earth nitrogen is present as N2, which is most stable for any homonuclear diatomic
molecule; highest bond dissociation energy; that’s wht inert so no life can utilise it;
nitrogen fixation means conversion of N2 into ammonia.

Industry uses by Haber-Bosch process, which uses drastic condition.

The ammonia synthesized is used in amino acids and protein synthesis. The enzymes
which does nitrogen fixation are called nitrogenase. Mainly 3 types:
1. Vanadium nitrogenase
2. Iron nitrogenase
3. Molybdinum nitrogenase.
Each nitrogenase is composed of tro mettaloproteins: A Fe protein and a MFe protein
(M= Mo, V & Fe).
Molybdinum nitrogenase is most active.
Molybdinum nitrogenase