Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.5.4.4 (adenosine deaminase)
 5,136 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Dextran-linked adenosine deaminase (EC 3.5.4.4) has been prepared. The polymer-linked enzyme possesses an optimal enzymatic activity of 27 units/mg immobilized protein (non-bound enzyme: 200 units/mg protein). Support-bound adenosine deaminase (4.5 microgram protein/mg dextran) shows an enhanced heat stability, a moderately increased Km, and a decreased V value compared to those of the free enzyme. The pH dependences of V and pKm values of dextran-linked adenosine deaminase show only two inflection points compared to three for the free enzyme, which are equivalent to the pK values of the enzyme. Since the missing third inflection point (pH 9.8) can be assigned to the pK value of the epsilon-amino group of lysyl residues, it can be concluded, that immobilization of adenosine deaminase on cyanogen-bromide-activated dextran took place via these lysyl residues. The remaining pK values found from the other inflection points are moderately shifted owing to the altered secondary structure. From the temperature dependence of the enzymatic activity, a 40% decrease of the activation energy of the support-bound enzyme was found, indicating diffusion controlled deamination. The immobilization of adenosine deaminase results in a fluorescence quenching of 80%, without shifting the ultraviolet maximum of the emission spectrum. As already shown for unmodified dextran, the matrix of polymer-linked adenosine deaminase is degradable by a bacterial endodextranase (EC 3.2.1.11).
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PMID:Adenosine deaminase covalently linked to soluble dextran. The effect of immobilization on thermodynamic and kinetic parameters. 617 33

Dextran-bound adenosine, inosine, and nebularine have been prepared by carbodiimide coupling of their 2',3'-O-(4-carboxyethyl-1-methylbutylidene) cyclic acetal derivatives to 6-aminohexyldextran or 12-aminododecanyldextran. The latter polymers were prepared by cyanogen-bromide activation of dextran T80 followed by reaction with 1,6-diaminohexane or 1,12-diaminododecane. A high CNBr concentration leads to high-molecular-weight material, probably due to cross-linking, accompanied by a decrease in the digestion velocity using endo-dextranase from Penicillium species (EC 3.2.1.11). The dextran-bound nucleosides, as well as the nucleoside 2',3'-O-(4-ethoxycarbonyl-1-methylbutylidene) acetal derivatives, were tested as substrates and inhibitors for adenosine deaminase. The Km of the adenosine acetal ester is identical to that of adenosine which shows that acetalation does not hinder complex formation. Since the maximum velocity of deamination is decreased fourfold, the modified substrate does not fit as well as the nucleoside. The polymer-bound acetals show a 3-8-fold increase of Km or Ki and unchanged V compared to the corresponding acetals while dextranase digestion of the support does not alter the kinetic data. This indicates that the length of the polysaccharide chain does not interfere either with the complex formation or with the catalytic activity of the modified substrate. Since the activation energies of the deamination reactions of adenosine, its acetal ester, and dextran-linked adenosine are all similar (29.8-32.3 kJ mol-1) it is concluded that no diffusion control of the enzymatic reaction results from the binding of the nucleoside acetals to dextran T80.
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PMID:Dextran-linked purine nucleosides as substrates and inhibitors of adenosine deaminase. 618 16