Saturday, April 14, 2012

Cysteine

3-phosphoglycerate family: Cysteine (C)

General features
R group: -(CH2)-SH
Molecular mass: 121.16 g/mol
R-group pKa: 8.5
Physiological roles
Cysteine has several structural and catalytic roles: protein structure (disulfide bond formation for stability when secreted), cysteine proteases (big contributor to virulence), antioxidant properties (glutathione), iron-sulfur complexes, PPP reactions (transketolase contains coenzyme B1/thiamine. The sulfur in the thiazole ring functional group, which is crucial for catalytic function, is derived from cysteine.)

Thiazole synthesis:
A good diagram may be found here:

Disulfide bond formation:
Disulfide bonds are highly useful for protein structure, particularly because unlike all the other sidechain interactions that contribute to secondary and tertiary structure, disulfide bridges are covalent linkages, not distance dependent electrostatic and van der Waal forces. Disulfide bonds are ideal for secreted proteins because the physically hold protein folds in position (obvious) but the bridge stability is pH dependent like the other noncovalent interactions. Reducing environments can begin to reduce (and break) disulfide bonds at pH 7, but relative rates vary for given solutions and microenvironments. For a ‘test-tube’ demonstration, look at the kinetic analysis of reductive potential of isolated proteins in solutions with various dithiols, DMH, BMS, DTT:

Cysteine protease activity: One mode of peptide bond cleavage.
The mechanism of a cysteine protease:

These proteases use a conserved histidine residue to aid in their peptide bond attack. Once the acyl-enzyme intermediate is formed (with one portion of the previously intact protein released with a new free amino group) the Histidine aids in the release of the second portion of the cleaved protein. The histidine will pull a proton from a ambient water to generate a reactive –OH, this will attack the ‘vulnerable’ carbonyl carbon of the thioacyl intermediate (pictured on the far right).

Cysteine proteases specific for host cell zymogens are particularly important virulence factors for many bacteria, including those of Staphylococcus aureus and Streptococcus pyogenes. For example, details on S. pyogenes’ SpeB protein can be found here: http://mic.sgmjournals.org/content/150/5/1559.full

Glutathione synthesis:
http://www.brenda-enzymes.org/Mol/reaction-popup.php4?id=119&type=I&displayType=marvin

After cysteine and glutamine have been linked via the gamma (amide-containing carbon of glutamine) an ATP is used to activate a glycine that forms a peptide linkage to with carboxyl group of cysteine.

Proposed antioxidant roles of glutathione (Pittman, Robinson, Poole, 2004) :

Generally speaking, glutathione is used in the cell as protection from free radical damage, using the sulfhydryl groups of adjacent cysteine residues of adjacent glutathione molecules.  A disulfide bridge can be formed from two previously reduced cysteine residues contributing two electrons to two free radicals.

In the figure above, the oxidized glutathione is generated a couple ways, by DsbC (a disulfide bond forming enzyme) and by other proteins that may or may not have radicals but use the reducing power of glutathione. The regeneration of reduced glutathione is not well-understood in bacteria.

Synthesis


Figure 3 from Lithgow, et al. (2004) helps define cyteine’s role in sulfur acquisition in Escherichia coli and Bacillus subtilis. Sulfur can be translocated into the cell in various forms by various transporters, but ultimately, when inorganic sulfur is incorporated into cysteine (left side of the figure), some intermediate, sulfur is added either in S2O3- form or S2- form to O-acetylserine (OAS) which is generated by adding an acetyl group  (COCH3) to the hydroxyl group of serine’s sidechain.
O-acetylserine shown here at chemspider http://www.chemspider.com/ImageView.aspx?id=184
The general reaction may be found here in scheme 1 http://www.sciencedirect.com/science/article/pii/S0003986104004552

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