The Mycobacteria were the first bacteria shown to have multiple chaperonins (Kong et al., 1993; Lund, 2001). In M. tuberculosis there are two chaperonin genes, one (cpn60.1)

in an operon with the cochaperonin gene cpn10 and the other (cpn60.2) elsewhere on the chromosome (Kong et al., 1993). The latter encodes Hsp65 and its nomenclature as cpn60.2 genes reflect its distinct non-operon-encoded genomic localisation. Surprisingly, however, deletion studies in Mycobacterium smegmatis, M. tuberculosis and Mycobacterium bovis BCG this website have shown that cpn60.2, and not cpn60.1, encodes the essential chaperonin, despite the latter being operon-encoded with cpn10 as in E. coli (Ojha et al., 2005; Hu et al., 2008; Wang et al., 2011). This has led to some debate about the functional equivalence of the mycobacterial cpn60 and Selleckchem Nutlin3a the groEL genes (Lund, 2009). This controversy has not been resolved by the conflicting results obtained from studies on the oligomerisation of recombinant products of the different cpn60 genes and the crystal structures of their gene products (Qamra & Mande, 2004; Qamra et al., 2004; Lund, 2009). More recently, Lund and colleagues have addressed the questions posed by the presence

of multiple Cpn60 proteins and their state of oligomerisation by undertaking a detailed genetic and biophysical characterisation of the chaperonins from M. tuberculosis and M. smegmatis (Fan et al., 2012). These studies present evidence supporting the evolution of novel function for the cpn60.1 genes and show that the cpn60.2-encoded proteins are highly likely to function as oligomers in vivo as they assemble into oligomers in the presence of high salt and nucleotides. They also show that Cpn60.2 from both M. tuberculosis and M. smegmatis

is able O-methylated flavonoid to replace GroEL in E. coli, when expressed with either the cochaperonin GroES or the cognate cochaperonin Cpn10. However neither Cpn60.1 nor Cpn60.3, a third chaperonin homologue found in M. smegmatis, was able to complement GroEL in E. coli. These studies also addressed the question of oligomerisation using a number of biophysical techniques and confirmed earlier structural studies showing that, under normal physiological conditions, the purified chaperonins are largely monomers or dimers (Qamra et al., 2004; Fan et al., 2012). However, as monomeric GroEL is nonfunctional (Hartl & Hayer-Hartl, 2002), they examined oligomer formation under a range of conditions and showed oligomerisation in the presence of high concentrations of ammonium salts and either ATP or ADP. Under these conditions, the ATPase activity of the chaperonins increased and the oligomers formed had molecular masses consistent with the typical GroEL tetra-decameric structure of a double ring with seven subunits each. Finally, they showed that substitution of the 22 amino acids at the N-terminus of cpn60.