Calcium complexes enzyme stabilization

Calcium-binding proteins, 6, 564, 572, 596 intestinal, 6, 576 structure, 6, 573 Calcium carbonate calcium deposition as, 6, 597 Calcium complexes acetylacetone, 2, 372 amides, 2,164 amino acids, 3, 33 arsine oxides, 3, 9 biology, 6, 549 bipyridyl, 3, 13 crown ethers, 3, 39 dimethylphthalate, 3, 16 enzyme stabilization, 6, 549 hydrates, 3, 7 ionophores, 3, 66 malonic acid, 2, 444 peptides, 3, 33 phosphines, 3, 9 phthalocyanines, 2,863 porphyrins, 2, 820 proteins, 2, 770 pyridine oxide, 3,9 Schiff bases, 3, 29 urea, 3, 9... 

It should be pointed out that the addition of substances, which could improve the biocompatibility of sol-gel processing and the functional characteristics of the silica matrix, is practiced rather widely. Polyethylene glycol) is one of such additives [110— 113]. Enzyme stabilization was favored by formation of polyelectrolyte complexes with polymers. For example, an increase in the lactate oxidase and glycolate oxidase activity and lifetime took place when they were combined with poly(N-vinylimida-zole) and poly(ethyleneimine), respectively, prior to their immobilization [87,114]. To improve the functional efficiency of entrapped horseradish peroxidase, a graft copolymer of polyvinylimidazole and polyvinylpyridine was added [115,116]. As shown in Refs. [117,118], the denaturation of calcium-binding proteins, cod III parvalbumin and oncomodulin, in the course of sol-gel processing could be decreased by complexation with calcium cations. 

The effect of calcium acetate in the reaction medium at higher concentrations of 4 was rather surprising as the enzymatic reaction proceeded without any problem at substrate loads as high as 3M (765 gL" ) with conversion values ranging from 42 to 48% after 24h (see Table 8.2, entries 1-4). Since the level of calcium acetate used in these experiments did not exceed 170 mM, which is well below a stoichiometric ratio between the carboxylate of 13 and Ca , a more complex mechanism that probably involves enzyme stabilization as well as com-plexation of product might be taking place. 

The use of alkali and alkaline earth group metal ions, especially those of sodium, potassium, magnesium, and calcium, for maintenance of electrolyte balance and for signaling and promotion of enzyme activity and protein function are not discussed in this text. Many of these ions, used for signaling purposes in the exciting area of neuroscience, are of great interest. In ribozymes, RNAs with catalytic activity, solvated magnesium ions stabilize complex secondary and tertiary molecular structure. Telomeres, sequences of DNA at the ends of chromosomes that are implicated in cell death or immortalization, require potassium ions for structural stabilization. 

The inverting GH 95 1,2-oc-L-fucosidase (EC 3.2.1.63) from Bifidobacterium bifi-dum in complex (PDB 2EAC) with 1-deoxy-L-fuconojirimycin (20) showed the inhibitor in C4 chair conformation, resembling L-fucose and the entrance to the active site was narrowed by the gate keeper loops.315 A calcium ion associated with the protein was not involved in the catalytic process but was considered to stabilize the enzyme. 

Titration curves of trypsin were obtained under a variety of conditions by Duke et al. (1952). The most noteworthy feature is a specific effect of calcium, which displaces the acid part of the titration curve to lower pH, and decreases the total number of groups which are titrated between pH 6 to 9. It is likely that the groups titrated between pH 6 and 9 in the absence of Ca " are a-amino groups, produced by self-digestion of the enzyme. The effect of Ca" " thus appears to result from a complex with the carboxyl groups of the protein, which stabilizes the anionic form of these groups so as to produce the displacement of the acid part of the titration curve. This complex is more resistant to self-digestion than the enzyme alone. 

Caldolysin is the trivial name of the serine proteinase from T. aquaticus strain T351 [284]. This enzyme did not have detectable esterase aetivity and hydrolysis of small peptides of less than four amino acids was not observed. The enzyme was highly stable in the presenee of ealcium ions. Caldolysin bound six calcium ions per molecule of enzyme, there being both high and low affinity binding sites [285]. Stability of apocaldolysin (i.e. caldolysin treated with EDTA to remove all calcium ions) was restored upon incubation with either calcium or lanthanide ions, the latter giving a lanthanide-caldolysin complex more stable than the native enzyme. Strontium ions were the only other divalent metal ions tested that could restore more than 50% activity. 

Perhaps the most significant biological effect of the lanthanide contraction results from the fact that the REE ions have an ionic radius similar to the calcium ion Ca ". Within the lanthanides the radius of the tripositive ions ranges from 8.5 X 10 to 1.15 X 10 nm. Calcium has an ionic radius of 9.9 x 10 nm. The extra positive charge on the REE, as compared to Ca, will, in most cases, contribute to the stability of complexes formed by lanthanides (Mikkelson 1976). This results in the inhibition of many Ca " -dependent enzymes by the REE (see Polya et al. 1987). It is this similarity to and potential to replace and compete with Ca " that has been the emphasis of most research into rare earths in biological systems. 

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