Beschreibung:
<jats:title>Abstract</jats:title><jats:p>An important question in protein folding is whether compact substructures or domains are autonomous units of folding and assembly. The protomer of the tetrameric D‐glyceraldehyde‐3‐phosphate dehydrogenase from the hyperthermophilic bacterium <jats:italic>Thermotoga maritima</jats:italic> has a complex coenzyme‐binding domain, in which residues 1‐146 form a compact substructure with the last 31 residues (313‐333). Here it is shown that the gene of a single‐chain protein can be expressed in <jats:italic>Escherichia coli</jats:italic> after deleting the 163 codons corresponding to the interspersed catalytic domain (150‐312). The purified gene product is a soluble, monomeric protein that binds both NAD<jats:sup>+</jats:sup> and NADH strongly and possesses the same unfolding transition induced by guanidinium chloride as the native tetramer. The autonomous folding of the coenzyme‐binding domain has interesting implications for the folding, assembly, function, and evolution of the native enzyme.</jats:p>