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Journal Article

Preparation of active enzyme samples for IR studies of Na+, K+-ATPase

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Grell,  Ernst
Department of Biophysical Chemistry, Max Planck Institute of Biophysics, Max Planck Society;

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Citation

Thoenges, D., Tscherp, C., Grell, E., & Barth, A. (2002). Preparation of active enzyme samples for IR studies of Na+, K+-ATPase. Biopolymers, 67(4-5), 271-274. doi:10.1002/bip.10108.


Cite as: https://hdl.handle.net/11858/00-001M-0000-0024-DCA7-5
Abstract
In the case of the integral membrane protein Na+/K+-ATPase, preparation of highly concentrated samples for IR difference spectroscopy often leads to inactivation of the enzyme. Therefore, we compared the activity of Na+/K+-ATPase using different techniques of sample preparation. The loss of activity can be minimized by cooling the sample to 10°C and by the addition of glycerol and dithiothreitol. The activity of Na+/K+-ATPase isolated from pig kidney is independent of the protein concentration whereas the enzyme from shark rectal gland is inactivated at concentrations above 1 μg/μL and is thus unsuitable for IR experiments.