Fluorescence Competition Assay for the Assessment of ATP Binding to an Isolated Domain of Na+, K+-ATPase

Title:

Fluorescence Competition Assay for the Assessment of ATP Binding to an Isolated Domain of Na+, K+-ATPase

Creator:

Kubala, M., Plášek, J., and Amler, E.

Identifier:

https://cdk.lib.cas.cz/client/handle/uuid:454e11ff-724d-4571-be39-3bc7960a8054
uuid:454e11ff-724d-4571-be39-3bc7960a8054

Subject:

Competition, Dissociation constant, Fluorescence, Enzyme-ligand interaction, and TNP-ATP

Type:

article, model:article, and TEXT

Description:

An equation allowing estimation of the dissociation constant for binding of a non-fluorescent ligand to the enzyme is presented that is based on the competitive replacement of the ligand by its fluorescent analog. We derived an explicit formula for the probe fluorescence intensity, which is suitable for nonlinear least-squares analysis. We used this formula to evaluate the binding of ATP to the large cytoplasmic loop of Na+,K+-ATPase. The estimated value of KD (6.2 ± 0.7 mM) is comparable with the results from other laboratories for similar constructs obtained by a different method.

Language:

English

Rights:

http://creativecommons.org/licenses/by-nc-sa/4.0/
policy:public

Source:

Physiological research | 2004 Volume:53 | Number:1

Harvested from:

CDK

Metadata only:

false