Tyrosine equilibria

Triple point The temperature and pressure at which the solid, liquid, and vapor phase of a substance can coexist in equilibrium, 233 Tryptophan, 622t Tyrosine, 622t... 

Owing to the fully reversible equilibrium nature of the aldol addition process, enzymes with low diastereoselectivity will typically lead to a thermodynamically controlled mixture of erythro/threo-isomers that are difficult to separate. The thermodynamic origin of poor threo/erythro selectivity has most recently been turned to an asset by the design of a diastereoselective dynamic kinetic resolution process by coupling of L-ThrA and a diastereoselective L-tyrosine decarboxylase (Figure 10.47)... 

In hemoglobin M, histidine F8 (His F8) has been replaced by tyrosine. The iron of HbM forms a tight ionic complex with the phenolate anion of tyrosine that stabilizes the Fc3 form. In a-chain hemoglobin M variants, the R-T equilibrium favors the T state. Oxygen affinity is reduced, and the Bohr effect is absent. P Ghain hemoglobin M variants exhibit R-T switching, and the Bohr effect is therefore present. 

A classical approach to driving the unfavorable equilibrium of an enzymatic process is to couple it to another, irreversible enzymatic process. Griengl and coworkers have applied this concept to asymmetric synthesis of 1,2-amino alcohols with a threonine aldolase [24] (Figure 6.7). While the equilibrium in threonine aldolase reactions typically does not favor the synthetic direction, and the bond formation leads to nearly equal amounts of two diastereomers, coupling the aldolase reaction with a selective tyrosine decarboxylase leads to irreversible formation of aryl amino alcohols in reasonable enantiomeric excess via a dynamic kinetic asymmetric transformation. A one-pot, two-enzyme asymmetric synthesis of amino alcohols, including noradrenaline and octopamine, from readily available starting materials was developed [25]. 

Ai-Stearoylamino acids and their methyl esters were synthesized from enantiomeric and racemic forms of tyrosine, serine, alanine, and tryptophan (Fig. 16). Analogs of these molecules were investigated initially over 30 years ago by Zeelen and Havinga, who found stereochemical differentiation in the monolayer HjA isotherms of these materials (Zeelen, 1956 Zeelen and Havinga, 1958). We have extended this study using more sensitive Langmuir balances, a wider array of dynamic and equilibrium techniques, and the A-stearoyl methyl esters of the amino acids (Harvey et al., 1989 Harvey and Arnett, 1989). 

The instability of these chiral monolayers may be a reflection of the relative stabilities of their bulk crystalline forms. When deposited on a clean water surface at 25°C, neither the racemic nor enantiomeric crystals of the tryptophan, tyrosine, or alanine methyl ester surfactants generate a detectable surface pressure, indicating that the most energetically favorable situation for the interfacial/crystal system is one in which the internal energy of the bulk crystal is lower than that of the film at the air-water interface. Only the racemic form of JV-stearoylserine methyl ester has a detectable equilibrium spreading pressure (2.6 0.3dyncm 1). Conversely, neither of its enantiomeric forms will spread spontaneously from the crystal at this temperature. 

RPTK activation. The activity of RPTKs is normally suppressed in their quiescent state. This suppression is due to the numerous loose and unstructured conformations of the activation loop (A loop) within the catalytic domain the majority of these conformations interfere with substrate and ATP binding. However, a subset of these conformations is amenable to binding of substrate and ATP, resulting in activation of the RPTKs. Phosphorylation of the tyrosine residue(s) in the A loop shifts the equilibrium towards the active conformation. Because of steric hindrance, PTK catalytic domains appear to be unable to autophosphorylate tyrosine residue(s) in the A loop within the same molecule rather frans-autophosphoryla-tion between two different catalytic domains is necessary for their activation. As a consequence, ligand-induced dimerization is an important step in the activation of RPTKs (Fig. 24-7). 

Interestingly, we have recently identified a mutation of a tyrosine in the third intracellular loop of the hDAT that causes a major alteration in the conformational equilibrium of the transport cycle, and thus as such is comparable to mutants on G protein-coupled receptors causing constitutive isomerization of the receptor to the active state (66). Most importantly, this conclusion is based on the observation that mutation of the tyrosine completely reverts the effect of Zn2+ at the endogenous Zn2+ binding site in the hDAT (50,51) from potent inhibition of transport to potent stimulation of transport (Fig. 6). In the absence of Zn2+, transport capacity is reduced to less than 1% of that observed for the wild-type, however, the presence of Zn2+ in only micromolar concentrations causes a close to 30-fold increase in uptake (66). Moreover, it is found that the apparent affinities for cocaine and several other inhibitors are substantially decreased, whereas the apparent affinities for substrates are markedly increased (66). Notably, the decrease in apparent cocaine affinity was around 150-fold and thus to date the most dramatic alteration in cocaine affinity reported upon mutation of a single residue in the monoamine transporters (66). 

Loland, C. J., Norregaard, L., Litman, T and Gether, U. (2002) Generation of an activating Zn(2+) switch in the dopamine transporter mutation of an intracellular tyrosine constitu-tively alters the conformational equilibrium of the transport cycle. Proc. Natl. Acad. Sci. USA 99,1683-1688. 

Levitzki A (2000) Protein Tyrosine Kinase Inhibitors as Therapeutic Agents. 211 1-15 Lewis, FD, Wasielewski MR (2004) Dynamics and Equilibrium for Single Step Hole Transport Processes in Duplex DNA. 236 45-65... 

Intramolecular interactions between two coordinated amino acids can influence the position of-cis trans equilibrium. These interactions can take the form of hydrophobic stacking interactions, as observed between the side chains of tyrosines in [Cu(L-TyrO)2J and [Pd(L-TyrO)2], or Coulombic attraction between oppositely charged side chains, as in [Cu(HisO)L] (L = Arg, Lys or ornithine).51 The optical configuration of the amino acids is of particular importance for these interactions. For amino acids of the same configuration trans geometry (15) is required, but for amino acids of opposite configuration cis geometry (14) is necessary around the metal centre. 

In the case of oligomeric proteins in which subunit contact regions have been revealed by X-ray crystallography34 353 or other methods described above,363 the equilibrium between oligomer and monomer can be changed by site-directed mutagenesis. For example, stable monomers of tyrosyl-tRNA synthetase were produced by a mutation of Phe-164 at the subunit interface to Asp, and it was revealed that the monomers are inactive and do not bind the substrate tyrosine.343 In the case of yeast triosephosphate isomerase, replacement of Asn-78 at the subunit interface did not cause dissociation of subunits under normal conditions.353 However, the stability of the enzyme was significantly lowered by the mutation, probably due to decreased subunit-subunit interaction.353... 

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