ELECTRON TRANSFER IN

BIOLOGY

Structure and Function of Metalloproteins in

Electron Transport Processes : Rubredoxin,
Cytochromes and Iron-Sulphur Proteins
 RUBREDOXIN
 Rubredoxins are a class of low-molecular-weight iron-
containing proteins found in sulphur-metabolizing bacteria.
 These are non-haem iron protein having one iron(III) active
centre.
 Sometimes rubredoxins are classified as iron-sulphur
proteins however, in contrast to iron-sulphur proteins,
rubredoxins do not contain inorganic sulphide.
 Like cytochromes, ferredoxins and Rieske proteins,
rubredoxins participate in electron transfer in biological
systems.
 STRUCTURE & FUNCTION OF RUBREDOXIN
 Rubredoxin active site contains an iron ion which is coordinated to
four sulphur conserved cysteine residues (non labile) forming a
distorted tetrahedron.
 This is sometimes denoted as a [1Fe-0S] or an Fe1S0 system, in
analogy to the nomenclature for iron-sulphur proteins.
 Rubredoxins perform one-electron transfer processes. The central iron
atom changes between the +2 and +3 oxidation states. In both
oxidation states, the metal remains high spin, which helps to minimize
structural changes. The reduction potential of a rubredoxin is typically
in the range +50 mV to -50 mV.
 This iron-sulphur protein is an electron carrier, and it is easy to
distinguish its metallic centre changes: the oxidized state is reddish
(due to a ligand metal charge transfer), while the reduced state is
colourless.
 Fe(III)
↑ ↑↑ ↑↑
EPR Active ( S = 5/2)
(Oxidised Form)

↓Gain of Electron

Fe(II)
↓↑ ↓ ↓ ↓ ↓
EPR Active ( S = 2)
(Reduced Form)
 CYTOCHROMES
 Cytochromes are proteins containing heme as a cofactor. They are
classified according to the type of heme and its mode of binding. Four
varieties are recognized, cytochromes a, cytochromes b, cytochromes c and
cytochrome d.
 In addition to the above classification  into four cytochrome classes, several
additional classifications such as cytochrome- o and cytochrome P450 can
be found in biochemical literature.
 Cytochrome function is linked to the reversible redox change
from ferrous(Fe(II)) to the ferric(Fe(III)) oxidation state of the iron found in
the heme core.
 Cytochromes were initially described in 1884 by MacMunn as respiratory
pigments. In the 1920s, Keilin rediscovered these respiratory pigments and
named them the cytochromes, or “cellular pigments.  He classified these
heme proteins on the basis of the position of their lowest energy absorption
band in their reduced state, as cytochromes a (605 nm), b (≈565 nm),
and c (550 nm).
 CYTOCHROMES
 Cytochrome is a protein that can transfer electrons with a chemical
group called a heme group. The heme groups of cytochrome are
similar to those of hemoglobin. Both have the same basic ring
structure called a porphyrin ring. 
 The heme group is a highly conjugated ring system surrounding an
iron ion. The iron in cytochromes usually exists in a ferrous (Fe2+) and
a ferric (Fe3+) state with a ferroxo (Fe4+) state found in catalytic
intermediates.
 Cytochromes are, thus, capable of performing electron transfer
reactions and catalysis by reduction or oxidation of their heme iron.
The cellular location of cytochromes depends on their function. They
can be found as globular proteins and membrane proteins.
 In the process of oxidative phosphorylation, a globular cytochrome c
protein is involved in the electron transfer. Photosystem II, the
first protein complex in the light-dependent reactions of
oxygenic photosynthesis, contains a cytochrome b subunit. Cyclo-
oxygenase 2, an enzyme involved in inflammation, is a cytochrome b
protein.
 CYTOCHROME P450
 It is hexa co-ordinated Fe(III) low spin heme iron protein.
 Four places are occupied by nitrogen atoms of porphyrin ring and 5th and 6th
position are occupied by water and sulphur cysteine residue respectively.
 The CYP450 enzymes are essential for the production of numerous agents
including cholesterol and steroids.
 CYP450 enzymes are so named because they are bound to membranes
within a cell (cyto) and contain a heme pigment (chrome and P) that
absorbs light at a wavelength of 450 nm when exposed to carbon
monoxide(soret band – blue to red).
 Cytochrome P450 enzymes are primarily found in liver cells but are also
located in cells throughout the body. Within cells, cytochrome P450
enzymes are located in a structure involved in protein processing and
transport (endoplasmic reticulum) and the energy-producing centers of cells
(mitochondria).
 The colour of cytochrome is due to Л to Л*.
 STRUCTURE AND FUNCTION
 It facilitates cleavage of O2.
 R-H + O2 + 2H + 2e- R–OH + H2O
 R–CHO R–COOH
 Converts alkene to epoxide.
 IRON-SULPHUR PROTEINS(FERREDOXIN)
 Ferredoxins are small proteins containing iron and sulfur atoms organized
as iron–sulfur clusters. These biological "capacitors" can accept or
discharge electrons, with the effect of a change in the oxidation state of the
iron atoms between +2 and +3. In this way, ferredoxin acts as an electron
transfer agent in biological redox reactions.
 Ferredoxins are present in all organisms, from bacteria to higher eukaryotes.
They are small and acidic proteins with prosthetic groups composed of iron
and sulphur atoms, which are referred to as an iron-sulphur cluster.
 The ferredoxins function as intracellular electron carriers with a low
reduction potential, and do not carry out classical enzyme functions.
 Ferredoxins are classified into several groups according to the types and
numbers of their Fe-S clusters.
 Even though each cluster contains more than one iron atom, each cluster
can carry only one extra electron. Thus ferredoxins that contain a single
cluster participate in single electron transfer reactions, while ferredoxins
that carry two clusters can transfer two electrons simultaneously.
 Fe2S2 or 2Fe-2S Ferridoxin
 They are proteins of around one hundred amino acids with four conserved
cysteine residues to which the 2Fe-2S cluster is ligated.
 Originally found in chloroplast membranes, has been termed 'chloroplast-
type' or 'plant-type', and includes ferredoxins from plants, algae,
rhodobacter and a toluene degrading pseudomonas.
 In 2Fe-2S, the irons are tetrahedrally coordinated by both inorganic
sulphurs and sulphurs provided by 4 conserved Cysteine residues.
 In chloroplasts, 2Fe-2S ferredoxins function as electron carriers in the
photosynthetic electron transport chain and as electron donors to various
cellular proteins.
 In oxidised form both Fe active centres are in +3 state and is diamagnetic
and ESR inactive whereas in reduced form one Fe centre is in +3 and other
in +2 state due to which it becomes paramagnetic and ESR active.
 When any two S-cysteine residues are changed by N-histidine group, the
compound so formed is called RIESKE protein.
STRUCTURE
Fe(III) Fe(III)
↑ ↑↑ ↑↑ ↓↓↓ ↓ ↓
EPR Inactive ( S = 0)
(Oxidised Form)

↓Gain of Electron

Fe(III) Fe(II)
↑ ↑↑ ↑↑ ↓↑ ↓ ↓ ↓ ↓
EPR Active ( S = 1/2)
(Reduced Form)
 Fe3S4 or 3Fe-4S Ferridoxin
 The [3Fe-4S] cluster has the shape of a cube with one vertex missing.
 In [4Fe-4S] ferredoxins the Fe ions are ligated by four L-cysteine residues
that occur in a -C-X2-C-X2-C-Xn- pattern. Omission of one of the Cys
residues from the conserved motif (usually the third) gives rise to the 3Fe-
4S ferredoxins.
 3Fe-4S cluster ha 3 Fe active sites, each Fe centre is coordinated with 3
labile Sulphur atoms and one Non-labile S-cysteine residue.
 All the three Fe centres are in +3 state in oxidised form and in reduced
form two Fe centres are in +3 state and one in +2 state. Thus both oxidised
and reduced form of 3Fe-4S cluster are paramagnetic and ESR active.
 These proteins have a distinctive red colour due to a sulfur to iron charge
transfer (LMCT).
STRUCTURE
Fe(III) Fe(III) Fe(III)
↑ ↑↑ ↑↑ ↓↓↓ ↓ ↓ ↑ ↑↑ ↑↑
EPR/ESR Active ( S = 5/2)
(Oxidised Form)

↓Gain of electron

Fe(III) Fe(III) Fe(II)

↑ ↑↑ ↑↑ ↓↓↓ ↓ ↓ ↑↓ ↑ ↑ ↑ ↑
EPR/ESR Active ( S = 2)
(Reduced Form)
 Fe4S4 or 4Fe-4S Ferredoxin
 The Fe4S4 cluster has a cubane-type structure which can be visualized as a
dimer of two Fe2S2 units. The sulfido groups in Fe4S4 bridge three iron
centers rather than the two in Fe2S2
  The Fe4S4 cluster exists in two oxidation levels, an oxidized form [Fe4S4]2+2 
and a reduced form [Fe4S4]+. The oxidized form is comprised of two Fe(III)
and two Fe(II) and the reduced form, of one Fe(III) and three Fe(II).
 Thus Oxidised form is diamagnetic and ESR inactive whereas reduced form
is paramagnetic and ESR active.
 In the oxidized state, [Fe4S4]2+2, electrons are delocalized over the cluster so
that the average oxidation state of each iron is effectively 2.5 whereas in
reduced state average oxidation state of each iron is 2.25.
 These are found in bacteria and called as bacterial-type ferredoxin.
 The [Fe4S4] ferredoxins may be further subdivided into low-potential
(bacterial-type) and high-potential (HiPIP) ferredoxins.
 The formal oxidation numbers of the iron ions can be [2Fe3+, 2Fe2+] or
[1Fe3+, 3Fe2+] in low-potential ferredoxins. The oxidation numbers of the
iron ions in high-potential ferredoxins can be [3Fe3+, 1Fe2+] or [2Fe3+, 2Fe2+].
STRUCTURE
Fe(III) Fe(III) Fe(II) Fe(II)
↑ ↑↑ ↑↑ ↓↓↓ ↓ ↓ ↑↓↑↑ ↑↑ ↓↑ ↓ ↓ ↓ ↓
EPR/ESR Inactive ( S = 0)
(Oxidised Form)

↓ Gain of electron

Fe(III) Fe(II) Fe(II) Fe(II)

↑ ↑↑ ↑↑↑ ↓↑ ↓ ↓ ↓ ↓ ↑ ↓ ↑ ↑ ↑ ↑ ↓↑ ↓ ↓ ↓ ↓
EPR/ESR Active ( S = 1/2)
(Reduced Form)