Mass spectrometric determination of association constants of adenylate kinase with two noncovalent inhibitors.
- Daniel JM Department of Chemistry, Swiss Federal Institute of Technology, ETH, CH-8093, Zürich, Switzerland.
- McCombie G
- Wendt S
- Zenobi R
- 2003-05-15
Published in:
- Journal of the American Society for Mass Spectrometry. - 2003
Adenylate Kinase
Animals
Chickens
Enzyme Inhibitors
Molecular Structure
Muscle, Skeletal
Protein Binding
Spectrometry, Mass, Electrospray Ionization
English
Noncovalent complexes between chicken muscle adenylate kinase and two inhibitors, P(1),P(4)-di(adenosine-5')tetraphosphate (Ap4A) and P(1),P(5)-di(adenosine-5') pentaphosphate (Ap5A), were investigated with electrospray ionization mass spectrometry under non-denaturing conditions. The nonconvalent nature and the specificity of the complexes are demonstrated with a number of control experiments. Titration experiments allowed the association constants for inhibitor binding to be determined. Problems with concentration dependent ion yields are circumvented by a data evaluation method that is insensitive to the overall ionization efficiency. The K(a) values found were 9.0 x 10(4) M(-1) (Ap4A) and 4.0 x 10(7) M(-1) (Ap5A), respectively, in very good agreement with available literature data.
- Language
-
- English
- Open access status
- bronze
- Identifiers
-
- DOI 10.1016/S1044-0305(03)00132-6
- PMID 12745213
- Persistent URL
- https://sonar.ch/global/documents/222286
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