Adaptive enzyme formation

In addition there is other evidence pointing to the fact that the same enzyme is involved in reactions with both D-fructose and L-arabinose. First, the relative rates of reaction with D-fructose and L-arabinose, respectively, remain constant after partial inactivation of the enzyme by heat. Second, the enzyme catalyzing both reactions is produced to a marked extent when sucrose is used as substrate for the growth of the organisms, but not when D-glucose or L-arabinose is used sucrose phos-phorylase is an adaptive enzyme. Third, on fractionation of the enzyme preparation with various concentrations of ammonium sulfate, the relative activities of the fractions are the same for both sugars. These observations indicate not only that the same enzyme is involved in both reactions but also that no additional enzyme is required for the formation of D-glucosyl-L-arabinose. 

They stated further that, the new adaptive enzyme catalyzing Reaction 3 appears to be similar to the malic enzyme of pigeon liver, although strictly DPN (instead of TPN)-specific. The coenzyme specificity explains the ready occurrence of Reaction 1. Therefore, the authors showed that exogenous NAD was required for the overall reaction (malic acid -> lactic acid), but because this activity was measured manometrically, they never demonstrated the formation of reduced NAD. Similarly, they did not attempt to show that pyruvic acid was the intermediate between L-malic acid and lactic acid. Instead, the formation of pyruvic acid was inferred from the NAD requirement and because the malic acid dissimilation activity remained constant during purification while the lactate dehydrogenase activity decreased (14). In fact, attempts to show any appreciable amounts of pyruvic acid intermediate failed (22). 

That hydroxylamine might not be an obligatory intermediate, or occur as a free intermediate, in the reduction of nitrite to ammonia is suggested by the properties of nitrite reductases of Azotobacter chroococcum and Escherichia coli. The former is an adaptive enzyme, the formation of which requires nitrate or nitrite in the culture (31,2). It is FAD-depen-dent and presumably contains metals and p-mercuribenzoate inhibitable... 

A very intensive study of the adaptive enzyme jS-galactosidase of E. colt has been made by Monod and Cohn and their associates. Their studies have been carried out with growing cells. In these organisms free amino acids are rapidly incorporated into proteins, and there is no evidence of preformed protein precursors. j8-Galactosidase formation is... 

Induced cells contain a mechanism for concentrating inducers within the cells, and this mechanism appears to play a part in the induction process. The formation of enzyme depends on the continued presence of inducer. The rate of enzyme synthesis with adequate amoimts of inducer is proportional to the growth of the bacteria. When the inducer is removed (by suspending the centrifuged bacteria in fresh medium), enzyme synthesis stops abruptly. The enzyme already formed, however, is stable, and persists unchanged for many generations. Sulfur-labeled amino acids have been used to demonstrate that the induced enzyme is formed directly from free amino acids, and that proteins already in the bacteria do not contribute amino acids to the new enzyme. In the absence of the inducer, the adaptive enzyme retains its label. Some properties of inducers were found in a study of penicillinase production by BadUus cereusJ With this system it was shown that in a brief exposure a small amount of penicillin is specifically bound within the cells, and is not hydrolyzed, but stimulates the production of several equivalents of penicillinase. 

Figure 6.8 Schematic diagram of the enzyme DsbA which catalyzes disulfide bond formation and rearrangement. The enzyme is folded into two domains, one domain comprising five a helices (green) and a second domain which has a structure similar to the disulfide-containing redox protein thioredoxin (violet). The N-terminal extension (blue) is not present in thioredoxin. (Adapted from J.L. Martin et al.. Nature 365 464-468, 1993.)...

Figure 6. Enzymes act as recycling catalysts in biochemical reactions. A substrate molecule binds (reversible) to the active site of an enzyme, forming an enzyme substrate complex. Upon binding, a series of conformational changes is induced that strengthens the binding (corresponding to the induced fit model of Koshland [148]) and leads to the formation of an enzyme product complex. To complete the cycle, the product is released, allowing the enzyme to bind further substrate molecules. (Adapted from Ref. 1). See color insert.

The following diagram, adapted from that first presented by Bennett et alC, describes a postulated pathway for evolution of a protein dimer from single-domain proteins. The scheme begins with the fusion of two singledomain polypeptides and proceeds through the evolution of interdomain contacts, and in the case of enzymes, development of an active site. These same interdomain contacts can also stabilize formation of a domain-swapped dimer which then undergoes further evolution into a present-day dimer. 

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