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Substrate Analogs Their Use in Limiting

6 Substrate Analogs Their Use In Limiting Secondary Reactions [Pg.86]

Enzymes that catalyze unwanted secondary reactions are to be avoided in HPLC analysis. When such secondary reactions make it difficult to quantitate [Pg.86]

Analogs can be used in another way. Consider the development of an assay procedure for adenosine kinase, the enzyme that catalyzes the primary reaction Ado + ATP — AMP + ADP. Problems will arise during the assay of this activity in crude extracts, since other enzymes may be present that can form AMP directly from ATP. [Pg.87]

Radiochemical analogs such as radiolabeled adenosine are ideal for solving this problem, because if the formation of radiolabeled AMP is monitored, it is possible to distinguish the AMP formed from adenosine from that formed from ATP, which, of course, would not be labeled. [Pg.87]

The concentration of the substrate and other components of the reaction mixture must be established before the reaction is initiated. Decisions regarding enzyme concentration and sampling intervals must be made empirically with some information about the amount of expected product formation. [Pg.87]




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In limitation

Limited use

Limiting substrate

Their Use

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