Molecular cloning, expression, purification and characterization of a recombinant FKBP family member from Pseudomonas aeruginosa.

Abstract

The FK-506 binding proteins (FKBPs) are a family of proteins with characterised molecular functions such as peptidylprolyl cis-trans isomerase activity and chaperone activity. However, the FKBP family member from Pseudomonas aeruginosa is currently uncharacterised. This study has successfully developed a recombinant method of overexpressing and purifying the FK-506 binding domain of P. aeruginosa (PaFKBD) in a SUMO-PaFKBD fusion protein state. The characterisation of SUMO-PaFKBD revealed peptidylprolyl cis-trans isomerase activity with catalytic efficiency of 1.01 x 105 s-1 M-1 and this activity is inhibited by FK-506 with an IC50 of 92.6nM. More studies need to be done to further reaffirm these results and to further characterise the protein.