Title: ATPase Activity Measurements Using Radiolabeled ATP
Abstract: ATP provides the energy that is essential for all P-type ATPases to actively transport their substrates against an existing gradient. This ATP hydrolysis can be measured using different methods. Here, we describe a method that uses radiolabeled [γ-32P]ATP, which is hydrolyzed by P-type ATPases to ADP and 32Pi. Activated charcoal is used to bind the excess of [γ-32P]ATP, which can be separated from the unbound 32Pi by centrifugation. With this method, a wide range (0.1 μM–10 mM) of ATP can be used. In addition, we also describe in detail how ATP hydrolysis is translated into ATPase activity.
Publication Year: 2016
Publication Date: 2016-01-01
Language: en
Type: article
Indexed In: ['crossref', 'pubmed']
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Cited By Count: 1
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