Abstract
Five spin labeled derivatives of a neurotoxin from cobra venom were analyzed by the earlier suggested method. The procedure was adjusted to the complex motional behaviour of the label. Each protein derivative carried covalently bound spin label on different lysine residues. In two derivatives, at positions Lys44 and Lys46, the labels were strongly mobile, whereas for other three derivatives modified at Lys15, Lys25 and Lys26 the label was less mobile with respect to the protein molecule, which made possible determination of the rotational correlation time of the protein molecule (2.8±0.3 ns). The rotational correlation time was in good agreement with the calculated value for the rigid sphere of the corresponding molecular weight. On the basis of the estimate of the anisotropic motion degree, it was found from the order parameter S that the label mobility increases in the following series of lysine residues: Lys26, Lys25, Lys15, Lys46, and Lys44. From the analysis of positions of outer wide peaks in ESR spectra obtained by varying temperature and viscosity of the medium, we determined the parameters for computer simulation. The theoretical and experimental spectra were found to be in good agreement.
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Abbreviations
- Τ :
-
rotational correlation time of the protein molecule
- Τ l :
-
rotational correlation time of the spin label
- 2A Z , 2A⊥:
-
the rigid limit distance between OWP and IWP, respectively, for ESR spectra of spin labeled proteins
- 2ā, 2ā⊥:
-
the averaged limit distance between the OWP and IWP correspondingly mobile spin label to respect of protein moiety with; Τ = ∞
- 2A∼',2A′⊥:
-
distance between OWP and IWP in the ESR spectra of spin labeled proteins for any T and η media
- SL:
-
spin label
- NT:
-
neurotoxin II from cobra venom
- NT-SL-Lys44:
-
neurotoxin spin labeled at Lys44 residue
- OWP:
-
outer wide peaks in the immobilized ESR spectra
- IWP:
-
inner wide peaks in the immobilized ESR spectra
- WL:
-
the residual linewidth
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Timofeev, V.P., Tsetlin, V.I. Analysis of mobility of protein side chains by spin label technique. Biophys. Struct. Mechanism 10, 93–108 (1983). https://doi.org/10.1007/BF00535545
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DOI: https://doi.org/10.1007/BF00535545