Tryptophan-activating enzyme and

The molecular interaction between the activating enzyme and the amino acid is not known, but it probably varies with the type of activating enzyme. Some enzymes (. g., the alanine-activating enzyme) are unaffected by paramercuric benzoate, but others, like the tryptophan-activating enzyme, appear to be SH enzymes the activity of which depends on the presence of free SH groups in the molecule. Potassium ions are known to activate the tyrosine enzyme. 

The existence of an acyl-adenylate bound to enzyme has been demonstrated in the case of tryptophan activation (344). Tryptophan was incubated with ATP in the presence of substrate quantities of purified tryptophan-activating enzyme. Adenyl-tryptophan was isolated and chemically characterized after its removal from the denatured enzyme. 

In animal studies, high levels of cortisol have been shown to induce (increase) the activity of the enzyme tryptophan 2,3-dioxygenase in the liver, thereby decreasing the bioavailability of tryptophan to the brain. It is interesting to note that low acute doses of a number of different antidepressants inhibit the activity of this enzyme and, as a result, increase brain tryptophan concentrations, thus stimulating 5-HT synthesis (Badawy and Evans, 1982). In this way a link between the two key monoamine neurotransmitters and the hormone may be seen namely, reduced brain NA activity leads to decreased inhibition of the HPA axis, while increased levels of cortisol reduce 5-HT activity in the brain. Activation of the HPA axis has also been shown to result in tissue atrophy, in particular of the limbic system s hippocampus, and a reduction in the levels of neurotrophic factors responsible for the maintenance and optimal function of brain neurons (Manji et al., 2001). In conclusion, manipulation of the HPA axis (Nemeroff, 2002) and stimulation of neurotrophic factor activity (Manji et al., 2001) might open up new avenues for the treatment of affective disorders. 

CNTs can be functionalized with protein via non-covalent bond (Li et al., 2005 Kim et al., 2003 Mitchell et al., 2002). For example, (beta-lactamase I, that can be immobilized inside or outside CNTs, doesn t change enzyme s activity (Vinuesa and Goodnow, 2002). Taq enzyme can attach to the outside of CNT, and doesn t change its activity (Cui et al., 2004). Peptide with Histidine and Tryptophan can have selective affinity for CNT(Guo et al., 1998). Monoclonal antibody can attach to SWNTs. Protein-modified CNTs can be used to improve its biocompatibility and biomolecular recognition capabilities (Um et al., 2006). For example, CNTs functionalized with PEG and Triton X-100 can prevent nonspecific binding of protein and CNTs. Biotin moiety is attached to the PEG chains Streptavidin can bind specifically with biotin-CNT (Shim et al., 2002). 

A previous study of the reaction of ozone with lysoz3mie dissolved in anhydrous formic acid gave rise to the conclusion that the only amino acid residues affected in the early stages of the reaction were the tryptophan residues 108 and 111 ( ). Conversion of these residues to N -formyl-kynurenine did not cause loss in enzyme activity. Imoto et al. (7) have pointed out that this result is anomalous since modifications of tryptophan 108 (e.g. with iodine) normally causes inactivation. 

Estimation of the Binding Site. Tryptophan-108 shows a specific reaction with iodine, distinguishing it from other tryptophan residues of lysoz3mie. When try-108 is selectively oxidized by iodine, lysozyme completely loses its activity. Nevertheless, the lysozyme still shows the ability to form an enzyme-substrate complex with CM-chitin. This observation contributes to the conclusion that try-62 is an essential binding site for a complex formation (13). All ozonized lysozymes formed strong complexes with CM-chitin and could only be eluted by 0.2N HA (Fig. 6). This further confirms that two tryptophan residues (108 and 111) are indispensible for the hydrolytic action of lysozyme, and that inactivation by ozone cannot be attributed to inhibition of substrate binding capability. 

Isolation of alkaline phosphatase from Escherichia coli in which 85% of the proline residues were replaced by 3,4-dehydro-proline affected the heat lability and ultraviolet spectrum of the protein but the important criteria of catalytic function such as the and were unaltered (12). Massive replacement of methionine by selenomethionine in the 0-galactosidase of E. coli also failed to influence the catalytic activity. Canavanine facilely replaced arginine in the alkaline phosphatase of this bacterium at least 13 and perhaps 20 to 22 arginyl residues were substituted. This replacement by canavanine caused subunit accumulation since the altered subunits did not dimerize to yield the active enzyme (21). Nevertheless, these workers stated "There was also formed, however, a significant amount of enzymatically active protein in which most arginine residues had been replaced by canavanine." An earlier study in which either 7-azatryptophan or tryptazan replaced tryptophan resulted in active protein comparable to the native enzyme (14). 

Tryptophan oxygenase is another iron-containing enzyme which, like catalase, is inhibited by pyrazole in vivo but not in vitro. Again, however, the metabolite 4-hydroxypyrazole is active in vitro and shows, in contrast to catalase, competitive inhibition with tryptophan for the enzyme (79MI10505). Adrenaline and other phenols are also inhibitors of this enzyme and in this case, therefore, the heterocyclic ring appears not to be essential for activity. 

Distribution of Tryptophan Biosynthetic Enzyme Activities on Different Proteins in Bacteria and Fungi... 

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