Disulfide isomerase activity of the dynamic, trimeric Proteus mirabilis ScsC protein is primed by the tandem immunoglobulin-fold domain of ScsB

作者：Furlong Emily J; Choudhury Hassanul G; Kurth Fabian; Duff Anthony P; Whitten Andrew E^*; Martin Jennifer L^*

来源：Journal of Biological Chemistry, 2018, 293(16): 5793-5805.

DOI：10.1074/jbc.RA118.001860

Correct disulfide bond formation is essential for proper folding of many proteins, including bacterial virulence factors. The suppressor of copper sensitivity (Scs) proteins have roles in dithiol/disulfide interchange and the bacterial response to copper stress. Encoded in a four-gene cassette (ScsABCD) present in many Gram-negative bacteria, the Scs proteins are enigmatic and poorly characterized. Here, we show that the periplasmic alpha-domain of the membrane protein ScsB in the Gram-negative bacterium Proteus mirabilis forms a redox relay with the soluble periplasmic protein PmScsC. We also found that the periplasmic alpha-domain is sufficient to activate the disulfide isomerase activity of PmScsC. The crystal structure of PmScsB alpha at a resolution of 1.54 angstrom revealed that it comprises two structurally similar immunoglobulin-like folds, one of which includes a putative redox-active site with the sequence CXXXC. We confirmed the importance of these cysteine residues for PmScsB alpha function, and in addition, we engineered cysteine variants that produced a stable complex between PmScsC and PmScsB alpha. Using smallangle X-ray and neutron scattering analyses with contrast variation, we determined a low-resolution structure of the PmScsC-PmScsB alpha complex. The structural model of this complex suggested that PmScsB alpha uses both of its immunoglobulinlike folds to interact with PmScsC and revealed that the highly dynamic PmScsC becomes ordered upon PmScsB alpha binding. These findings add to our understanding of the poorly characterized Scs proteins.

• 出版日期2018-4-20