Structure and dynamics of the acidic compact state of apomyoglobin by frequency-domain fluorometry

Eur J Biochem. 1993 Nov 15;218(1):213-9. doi: 10.1111/j.1432-1033.1993.tb18367.x.

Abstract

The conformational dynamic properties of tuna apomyoglobin, a single tryptophan-containing protein, in the acidic compact state, as well as in the native and in the fully unfolded state, have been explored by frequency-domain fluorometry. Apomyoglobin at acidic pH in the presence of high salt concentration displays bimodal tryptophanyl lifetime distributions which may be related to the simultaneous presence of different populations of structural states (compact and fully unfolded states). The tryptophanyl anisotropy decay indicated that the acidic compact state displays at least two rotational correlational times, suggesting that this state possesses a complex geometrical organization. 1-Anilino-8-naphthalene sulfonate (ANS), bound both to native and compact protein forms, shows broad unimodal lifetime distributions. The small time dependence of the ANS emission spectra indicated that the solvent dipolar reorganization are either absent or they occur on a time scale much shorter than the lifetime of the excited ANS molecule bound to apomyoglobin. The anisotropy decay data relative to the extrinsic fluorophore (ANS) are consistent with the presence of a single rotational correlation time for both native (12.1 ns) and compact (6.2 ns) states.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Anilino Naphthalenesulfonates
  • Animals
  • Apoproteins / chemistry*
  • Fluorescence Polarization
  • Fluorescent Dyes
  • Hydrogen-Ion Concentration
  • Myoglobin / chemistry*
  • Osmolar Concentration
  • Protein Conformation
  • Tuna

Substances

  • Anilino Naphthalenesulfonates
  • Apoproteins
  • Fluorescent Dyes
  • Myoglobin
  • apomyoglobin
  • 1-anilino-8-naphthalenesulfonate