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Use of Helper Enzymes for ADP Removal in Infrared Spectroscopic Experiments: Application to Ca^sup 2+^-ATPase
- Article from:
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Biophysical Journal
- Article date:
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May 1, 2005
- Author:
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Copyright informationCopyright Biophysical Society May 2005. Provided by ProQuest LLC. (Hide copyright information)
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ABSTRACT
Adenylate kinase (AdK) and apyrase were employed as helper enzymes to remove ADP in infrared spectroscopic experiments that study the sarcoplasmic reticulum Ca^sup 2+^-ATPase. The infrared absorbance changes of their enzymatic reactions were characterized and used to monitor enzyme activity. AdK transforms ADP to ATP and AMP, whereas apyrase consumes ATP and ADP to generate AMP and inorganic phosphate. The benefits of using them as helper enzymes are severalfold: i), both remove ADP generated after ATP hydrolysis by ATPase, which enables repeat of ATP-release experiments several times with the same sample without interference by ADP; ii), AdK helps maintain the presence of ATP for ...
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