User:Estelle Blochouse/ Sandbox 1497

Copper is one of the most important metallic cofactor involved in enzyme catalysed reactions. In living organisms, its function is related to the redox properties of copper. However it is toxic at all concentration, therefore it needs to be strictly controlled by molecular mechanisms.^[1]

Function

Multicopper oxidases are enzymes involved in copper homeostasis. Indeed, free copper present in a cell (so, not bounded to a protein) is harmful and can cause cellular damage. It needs to be regulated.

Multicopper oxidase acts probably for the detoxification of copper present in the periplasmic space. It oxidizes the Cu+ into Cu2+ and prevents its uptake by the cytoplasm. It also possesses a phenoloxidase and a ferroxidase activity which can be involved in the prevention of oxidative damage.^[2]

Multicopper oxidase might also be involved in the regulation of metal transport.

Multicopper oxidases are able to oxidise their substrates thanks to their particular structure. They own four copper ions (Cu1001, Cu1002, Cu1003 and C1004) dispatched between two important areas : the mononuclear copper center (Cu1001) and the trinuclear copper center (Cu1002, Cu1003 and Cu1004).

They accept an electron in the and transfer it to the trinuclear copper center which binds to a molecule of dioxygen. The dioxygen receives four electrons from this transfer, therefore oxidation of the four copper ions allows reduction of dioxygen into water. It is transformed into two molecules of water.^[3] Three differents copper centres exist that can be differentiated spectroscopically: Type 1 or blue ()(Cu1001), type 2 or normal (Cu1004) and type 3 or coupled binuclear (Cu1002 and Cu1003).^[4][5]

Mechanism

Multicopper oxidase catalyzes the oxidation of different substrates by reducing O2 into H2O without releasing activated oxygen species (H2O2).

Multicopper oxidase contain 3 types of copper ions involved in the transfer of electrons from the substrate to the dioxygen. The first type of copper, type 1, (Cu1001) mediate the electron transfer from the substrate to the other coppers. The three other copper are called the trinuclear copper center. It is the key element for the oxygen reduction. It contains a type 2 copper (Cu1002) and 2 type 3 copper ions, binuclear ions, (Cu1003 and Cu1004). The final electron acceptor, O2, is bound to this last type of copper and is reduced into two molecules of water.^[6]

Disease

If the amino acids 500 and 501 are mutated from CH to SR, the residual activity and loss of resistance to copper. E. Coli die.

Structural highlights

Multicopper Oxidase CueO 4e9s is a protein containing one chain with sequence from Ecoli. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance.
Ligands:	Figure 1: Acetate ion.[7] ,
Related:	4e9p, 4e9q, 4e9r, 4e9t
Gene:	cueO, yacK, b0123, JW0119 (ECOLI)
Function:	binding of a metal ion
Process:	oxidation-reduction
Position:	bound at the outer membrane in the periplasmic space
Chain:	A
Sequence domain:	Cupredoxin, Multicopper oxidase type 1, type 2, type 3 and a copper-binding site
Number of amino acids:	489
Production and extraction:	Escherichia Coli
Initial gene:	sequence from amino acid 29 to amino acid 516 from P36649
Specific region:	contain a methionine rich region (aa355 to aa400) that can be important for copper tolerance in bacteria
Resources:	FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT, EMBL-EBI

In the chain, 5 ligands are present: an acetate ion (C2H3O2) and 4 copper ions.
Figure 2: Copper 1001[8]	Cu1001: The copper ion is bound thanks to 3 metal protein interactions with H443, H505 and C500, structure stabilised by L502, M510 by hydrogen bonds
Figure 3: Copper 1002[9]	Cu1002: 2 atoms of Cu, bonds twice by metal interaction (2x3) with 3 histidine : H501, H103, H141 stabilised by hydrophobic contact with W139
Figure 4: Copper 1003[10]	Cu1003: 2 CU bond with 3histidine : H499, H143, H448 by 2 metal interactions. h448 is stabilised by Pi interactions with [CU]1004 and H101
Figure 5: Copper 1004[11]	Cu1004: one single atome of CU bonds once by metal interactions with 2 H ( and one ACT[1005]) : H101 and H446 stabilised by Pi interactions with H103 and H448. [CU]1004 has Pi interactions with H103 and H448. H113 and H446 Pi interactions
Figure 6: Acetate ion 1005[12]	ACT[1005]: is linked by hydrogen bonds to G104 and G449

Cu1002, Cu1003, Cu1004 and ACT[1005] are near in the space, the 3 CU form a triangle.

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