Neutral protein stability

DAAO is one of the most extensively studied flavoprotein oxidases. The homodimeric enzyme catalyzes the strictly stere-ospecihc oxidative deamination of neutral and hydrophobic D-amino acids to give a-keto acids and ammonia (Fig. 3a). In the reductive half-reaction the D-amino acid substrate is converted to the imino acid product via hydride transfer (21). During the oxidative half-reaction, the imino acid is released and hydrolyzed. Mammalian and yeast DAAO share the same catalytic mechanism, but they differ in kinetic mechanism, catalytic efficiency, substrate specificity, and protein stability. The dimeric structures of the mammalian enzymes show a head-to-head mode of monomer-monomer interaction, which is different from the head-to-tail mode of dimerization observed in Rhodotorula gracilis DAAO (20). Benzoate is a potent competitive inhibitor of mammalian DAAO. Binding of this ligand strengthens the apoenzyme-flavin interaction and increases the conformational stability of the porcine enzyme. 

Although neutral methanol and ammonia are more stable in vacuo than their ions, the reaction field is capable of inverting this gap. At 3.0A as the spherical cavity radius, the diionic form becomes more stable. The tetrahedral substrate can approach the dyad to a shorter distance than the planar substrate. The repulsive barrier occurs at distances shorter than 2.5A for the planar, but only at 2.0A for the tetrahedral. The tetrahedral substrate is more stabilized by the reaction field effect than the planar substrate, due to an increase in the in-vacuo dipole moment of the tetrahedral. The reaction field is supposed to mimic the protein surrounding, and it is proposed that the protein stabilizes the diionic form even though the simulation of the reaction field is not sufficient to obtain a realistic interpretation. This study indicates a tendency to tetrahedralization of the model substrate at distances characteristic of the Michaelis-Menten complex formation. The authors believe that this must affect intermolecular interactions of large substrates. 

Rangsansarid, J. and Fukada, K., Factors affecting the stabihty of O/W emulsion in BSA solution Stabilization by electrically neutral protein at high ionic strength,... 

Although protein stability can be improved by neutralizing the detrimental acidic microenvironment, and without adjustment of this environment it may be difficult to stabilize an acid-labile protein, most proteins do require additional... 

The conformational stability of biomolecules is greatly dependent on the solvent species. It is also affected by coexisting solutes such as salts (e.g., NaCl). The salt effects [47, 48, 49] on the solubility and the conformational stability of proteins in aqueous solutions are experimentally known to follow the order called the Hofmeister series. The series for anions is [S04 > CHsCOO" > Cl > Br > NOa" > CIOJ > 1 > CNS ], and that for cations is [(CH3)4N+ > NH > Rb+,K+,Na+, Cs+ > Li+ > Mg + > Ca + > Ba +j. In each of these series, the species to the left decrease the solubility of proteins and stabilize their native structures. The species to the right, on the contrary, increase the solubility and cause destabilization of the native structures. Though the Hofmeister series is not valid for acidic and basic proteins [50, 51], it is generally applicable to neutral proteins. The series, except for divalent cations, is also applicable to the other neutral substances such as benzene [52]. That is, the effects of monovalent ions on the solubility of various neutral substances follow the Hofmeister series. The microscopic mechanisms of these experimentally known properties, however, have not been elucidated yet. 

Similar to the behavior of Nafion, poly(ester sulphonic acid) ionomers selectively exclude anionic species and large particles, but cations and neutral molecules are permeable. Of the three Eastman Kodak AQ 29, AQ 38, and AQ 55 polymers, AQ 55 (see the polymer backbone in Fig. 11.12) is the most studied and most applied in biosensor preparation. Wang and coworkers described have features of this polymer as an electrode material, including strong affinity toward hydrophobic counterions, prevention of electrode fouling from proteins, stability of the polymer film on the electrode, ability to preconcentrate catalysts in the film, and lowering the overpotential of many species difficult to oxidize or reduce. Several workers also showed that this poly(ester sulphonic acid) polymer is very stable as an organic phase electrode material. ... 

Why should the cores of most globular and membrane proteins consist almost entirely of a-helices and /3-sheets The reason is that the highly polar N—H and C=0 moieties of the peptide backbone must be neutralized in the hydrophobic core of the protein. The extensively H-bonded nature of a-helices and /3-sheets is ideal for this purpose, and these structures effectively stabilize the polar groups of the peptide backbone in the protein core. 

What about tertiary structure Why does any protein adopt the shape it does The forces that determine the tertiary structure of a protein are the same forces that act on ail molecules, regardless of size, to provide maximum stability. Particularly important are the hydrophilic (water-loving Section 2.13) interactions of the polar side chains on acidic or basic amino acids. Those acidic or basic amino acids with charged side chains tend to congregate on the exterior of the protein, where they can be solvated by water. Those amino acids with neutral, nonpolar side chains tend to congregate on the hydrocarbon-like interior of a protein molecule, away from the aqueous medium. 

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