One of the main functions of the Tat pathway in bacteria is to translocate prefolded metal-cofactor containing redox enzymes that are assembled in the anaerobic cytoplasm before translocation can occur. E. coli Tat substrates include enzymes that bind copper, molybdenum or Fe-S clusters and the folding of these substrates and the assembly of the metal cofactors into the apo-proteins must be somehow coordinated with the translocation process to ensure that proteins that are not properly assembled are not translocated prematurely

(Jack et al., 2004). This quality control mechanism is only just starting to be unravelled but several substrates Torin 1 chemical structure appear to have dedicated cytosolic chaperones that bind to the signal peptides of Tat substrates to prevent premature interactions with

the Tat machinery. Good examples of this from E. coli include the chaperones DmsD and TorD that bind specifically to signal peptides of the molybdenum-containing Tat substrates DmsA and TorA respectively (Ray et al., 2003; Jack et al., 2004; Hatzixanthis et al., 2005; Genest et al., 2006). The presence of similar chaperones in cyanobacteria selleck has yet to be demonstrated. An important study has recently discovered a central role for the Tat pathway in preventing the aberrant binding of metal ions by apo-proteins in Synechocystis (Tottey et al., 2008). Different metal ions have different binding affinities for different proteins, but the preference of a protein for a particular divalent metal ion usually follows the Irving-Williams series (Irving & Williams, 1948), Mn2+ < Fe2+ < Co2+ < Ni2+ < Cu2+ > Zn2+, although this

order can be influenced by steric effects imposed by proteins as well as by kinetics. When the Tat substrate MncA folds in the cytoplasm of Synechocystis, the apo-protein binds a manganese ion rather than a metal ion with a higher binding affinity, such as copper or zinc (Tottey et al., 2008). This is shown schematically in Fig. 2. In contrast, when MncA folds in periplasmic extracts, it binds more competitive zinc ions (Tottey et al., 2008). The bacterial cytoplasm is thought to contain essentially no free zinc or copper with all of these metal ions tightly bound to other bio-molecules (Rae et al., 1999; Outten & O’Halloran, Prostatic acid phosphatase 2001; Changela et al., 2003). In contrast, the cytoplasm is likely to contain free manganese at concentrations in the micromolar range (Helmann, 2007) allowing a kinetically favourable interaction between manganese and apo-MncA. Once assembled, folded and translocated to the periplasm by the Tat pathway, the much higher concentration of free copper and zinc within this compartment is unable to displace the bound manganese because it is deeply buried within the folded protein (Tottey et al., 2008). Metal ions are thought to diffuse freely into the periplasm through porins in the outer membrane, and the concentrations of metal ions within the periplasmic space are hence more dependent on the prevailing environmental concentrations.