Protein disulfide-isomerase

Protein disulfide isomerase or PDI is an enzyme in the endoplasmic reticulum in eukaryotes that catalyzes the formation and breakage of disulfide bonds between cysteine residues within proteins as they fold.^[1][2][3] This allows proteins to quickly find the correct arrangement of disulfide bonds in their fully folded state, and therefore the enzyme acts to catalyze protein folding.

Function

Protein folding

PDI contains four thioredoxin-like domains, two of which retain the canonical CXXC motif. The reduced (dithiol) form of PDI is able to catalyse a reduction of mispaired thiol residues of a particular substrate, acting as an isomerase.^[4] Therefore, PDI is capable of catalyzing the post-translational modification disulfide exchange. Such exchange reactions can occur intramolecularly, leading to the rearrangement of disulfide bonds in a single protein.^[5][5]

Another major function of PDI relates to its activity as a chaperone; i.e., it aids wrongly folded proteins to reach a correctly folded state without the aid of enzymatic disulfide shuffling.

Oxidized PDI can catalyze the formation of a disulfide bridge. This reduces PDI, which is re-oxidized by a protein called Ero1.

Redox signaling

In the chloroplasts of the unicellular algae Chlamydomonas reinhardtii the PDI RB60 serves as a redox sensor component of an mRNA-binding protein complex implicated in the photo-regulation of the translation of psbA, the RNA encoding for the photosystem II core protein D1. PDI has also been suggested to play a role in the formation of regulatory disulfide bonds in chloroplasts.^[6]

Other functions

PDI helps load antigenic peptides into MHC class I molecules. These molecules (MHC I) are related to the peptide presentation by antigen-presenting cells in the immune response.

PDI has been found to be involved in the breaking of bonds on the HIV gp120 protein during HIV infection of CD4 positive cells, and is required for HIV infection of lymphocytes and monocytes.^[7] Some studies have shown it to be available for HIV infection on the surface of the cell clustered around the CD4 protein. Yet conflicting studies have shown that it is not available on the cell surface, but instead is found in significant amounts in the blood plasma.

PDI is critical for thrombus formation.^[8]

Assays used for PDI activity

Insulin Turbidity Assay: PDI breaks the two disulfide bonds between two insulin (a and b) chains that results in precipitation of b chain. This precipitation can be monitored at 650 nm, which is indirectly used to monitor PDI activity.^[9] Sensitivity of this assay is in micromolar range.

ScRNase assay: PDI converts scrambled (inactive) RNase into native (active) RNase that further acts on its substrate.^[10] The sensitivity is in micromolar range.

Di-E-GSSG assay: This is the fluorometric assay that can detect picomolar quantities of PDI and therefore is the most sensitive assay to date for detecting PDI activity.^[11] Di-E-GSSG has two eosin molecules attached to oxidized glutathione (GSSG). The proximity of eosin molecules leads to the quenching of its fluorescence. However, upon breakage of disulfide bond by PDI, fluorescence increases 70-fold.

Inhibition

Due to the role of PDI in a number of disease states, small molecule inhibitors of PDI have been developed. These molecules can either target the active site of PDI irreversibly ^[12] or reversibly.^[13]

It has been shown that PDI activity is inhibited by red wine and grape juice, which could be the explanation for the French Paradox.^[14]

Members

Human genes encoding Protein disulfide isomerases include:^[3][15][16]

References

External links

• Protein Disulfide-Isomerase at the US National Library of Medicine Medical Subject Headings (MeSH)