Electrooptical measurements demonstrate a large permanent dipole moment associated with acetylcholinesterase.
Acetylcholinesterase (AChE) from krait (Bungarus fasciatus) venom is a soluble, nonamphiphilic monomer of 72 kDa. This snake venom AChE has been analyzed by measurements of the stationary and the transient electric dichroism at different field strengths. The stationary values of the dichroism are consistent with the orientation function for permanent dipoles and are not consistent with the orientation function for induced dipoles. The permanent dipole moment obtained by least-squares fits for a buffer containing 5 mM MES is 1000 D, after correction for the internal directing field, assuming a spherical shape of the protein. The dipole moment decreases with increasing buffer concentration to 880 D at 10 mM MES and 770 D at 20 mM MES. The dichroism decay time constant is 90 ns (+/- 10%) which is clearly larger than the value expected from the size/shape of the protein and indicates contributions from sugar residues attached to the protein. The dichroism rise times observed at low field strengths are larger than the decay times and, thus, support the assignment of a permanent dipole moment, although it has not been possible to approach the limit where the energy of the dipole in the electric field is sufficiently low compared to kT. The experimental value of the permanent dipole moment is similar to that calculated for a model structure of Bungarus fasciatus AChE, which has been constructed from its amino and acid sequence, in analogy to the crystal structure of AChE from Torpedo californica.
Porschke
D
D
Max-Planck-Institut für biophysikalische Chemie, Göttingen, Germany. dpoersc@gwdg.de
Créminon
C
C
Cousin
X
X
Bon
C
C
Sussman
J
J
Silman
I
I
D000110:Acetylcholinesterase / Q000737:chemistry*; D000818:Animals; D055592:Biophysical Phenomena; D001703:Biophysics; D017838:Bungarus; D004563:Electrochemistry; D008958:Models, Molecular; D055095:Optics and Photonics; D011487:Protein Conformation; D014101:Torpedo
D000110:Acetylcholinesterase