Absorption and fluorescence spectroscopic studies of the Ca2(+)-dependent lipid binding protein p36: the annexin repeat as the Ca2+ binding site

Biochemistry. 1990 Jul 31;29(30):7004-11. doi: 10.1021/bi00482a008.

Abstract

The existence of a single tryptophan residue in the protein p36, a member of a recently characterized family of Ca2+ binding proteins called annexins, is exploited to provide unique spectroscopic information on the annexin repeat motif and its role in Ca2+ binding. The differences in ultraviolet absorption and fluorescence excitation upon Ca2+ binding are interpreted solely in terms of this tryptophan, which, in view of the pronounced blue-shifts and the presence of vibronic structure, seems to reside in a highly nonpolar environment. The fluorescence emission from the protein is correspondingly blue-shifted, and it is found to transfer energy in resonance with Tb3+ absorption lines in the near-ultraviolet. This effect allows us to locate the Tb3+ and, by implication, the Ca2+ binding site to within ca. 8 A of the tryptophan residue.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Annexins
  • Binding Sites
  • Calcium / metabolism
  • Calcium-Binding Proteins / chemistry*
  • Calcium-Binding Proteins / metabolism
  • Chickens
  • Humans
  • In Vitro Techniques
  • Spectrometry, Fluorescence
  • Spectrophotometry, Ultraviolet
  • Swine
  • Terbium / metabolism
  • Tryptophan / chemistry

Substances

  • Annexins
  • Calcium-Binding Proteins
  • Terbium
  • Tryptophan
  • Calcium