Structural and functional characterization of TesB fromYersinia pestisreveals a unique octameric arrangement of hotdog domains

Abstract

Structural and functional characterization of the TesB thioesterase from Y. pestis reveals unique elements within the protomer and quaternary arrangements of hotdog domains, presenting distinguishing features of this enzyme family.

Structure, function, and regulation of thioesterases

“…New insights into a conserved and unique quaternary structure within TE4 family members was reported in 2015 [17]. An octamer of hotdog domains (or tetramer of the double hotdog domain protomers) was identified in the TesB from Y. pestis (PDB 4QFW) and confirmed using a range of biophysical and structural approaches including analytical ultracentrifugation, size exclusion chromatography, small angle X-ray scattering data, mutagenesis, and X-ray crystallography.…”

“…Members of the TE4 thioesterase family, also known as TesB, acyl-CoA thioesterase II, and ACOT8 thioesterases, exhibit substrate specificity for short-to long-chain acyl-CoA, palmitoyl-CoA, and choloyl-CoA substrates [17][18][19]. TE4 thioesterases contain a conserved double hotdog fold, and structures have been solved from both prokaryotes E. coli (PDB 1C8U; [20]), Yersinia pestis (PDB 4QFW; [17]), Mycobacterium avium (PDB 3RD7; [21]), Mycobacterium. marinum (PDB 3U0A; [21]), M. avium subsp.…”

“…Within related hotdog families several crystal structures have shown only half of the domains contain CoA, despite all domains containing equivalent active site configurations (PDBs: 4MOB, 4MOC [43], 3B7K, 4ZV3). Moreover, other single hotdog domain TE family members also exhibit CoA bound at only a fraction of the sites (PDBs 4R4U [17], 4ZRB [64], 5BYU, 5KL9). Substrate-induced structural rearrangements have been described in other thioesterase families [17,64], although the exact functional role for half-of-sites activity from prokaryotes through to eukaryotes remains to be established.…”

“…Moreover, other single hotdog domain TE family members also exhibit CoA bound at only a fraction of the sites (PDBs 4R4U [17], 4ZRB [64], 5BYU, 5KL9). Substrate-induced structural rearrangements have been described in other thioesterase families [17,64], although the exact functional role for half-of-sites activity from prokaryotes through to eukaryotes remains to be established. A half-of-sites reactivity has also been described more broadly, with a large number of enzymes including aldehyde J o u r n a l P r e -p r o o f dehydrogenase [65], thymidylate synthase [66], aspartate receptors [67] and bacterial adenylyltransferase [68], all displaying a half-of-sites activity.…”

Characterisation of four hotdog-fold thioesterases for their implementation in a novel organic acid production system

“…monomer, dimer and tretramer) of RpaL was unknown. Previous investigations looking into the biophysical properties of members of the TE4 family (TesB-like subfamily) containing a double hotdog domain have shown complex tetrameric structures to be formed (Pidugu et al 2009;Swarbrick et al 2015). SEC was used to determine the native size of RpaL, which showed a single protein elution corresponding to a biologically active size of~36 kDa, consistent with a monomer.…”