Institut for Biomedicin

Mikael Esmann

Vinyl ketone reagents for covalent protein modification. Nitroxide derivatives suited to rotational diffusion studies by saturation transfer electron spin resonance, using membrane-bound Na,K-ATPase as an example

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  • Mikael Esmann
  • H O Hankovszky
  • ,
  • K Hideg
  • ,
  • J A Pedersen, Danmark
  • D Marsh
  • Institut for Fysiologi og Biofysik
The reactivity of a series of substituted vinyl ketone nitroxides with an integral membrane protein, the Na,K-ATPase, is described. Increasing the electrophilicity of the conjugated double bond enhances reactivity markedly, with some spin labels showing higher reactivity than the conventionally used maleimide derivatives. The spectroscopic characteristics of the spin-labeled protein are also better suited for motional analysis by the saturation transfer electron spin resonance (STESR) method than with previous labeling procedures. The rotational correlation time, deduced from STESR experiments, is in the same range (100-300 microseconds) irrespective of the vinyl ketone derivative used, and the rotational mobility corresponds to an (alpha beta)2 or higher oligomer of the membrane-bound Na,K-ATPase.
OriginalsprogEngelsk
TidsskriftAnalytical Biochemistry
Vol/bind189
Nummer2
Sider (fra-til)274-282
Antal sider9
ISSN0003-2697
StatusUdgivet - 1 sep. 1990

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