Calcium- and myristoyl-dependent subcellular localization of the neuronal calcium-binding protein VILIP in transfected PC12 cells

Neurosci Lett. 1997 Apr 4;225(2):126-8. doi: 10.1016/s0304-3940(97)00201-2.

Abstract

Wild-type neuronal calcium-binding protein VILIP (visinin-like protein), and a myristoylation mutant of VILIP which lacks the consensus sequence for N-terminal myristoylation, have been stably transfected in PC12 cells. Immunocytochemical studies of VILIP-transfected PC12 cells have revealed the wild-type VILIP is strongly concentrated at the cell membrane, particularly at cell-cell contact sites, but is also distributed throughout the cytosol at moderate levels. In contrast, myristoylation-mutant VILIP shows a more even distribution, with significantly less association at cell-cell contact sites. Western blot analysis of subcellular fractions has shown that wild-type VILIP associates in a calcium-dependent manner with membrane fractions, whereas the myristoylation mutant only weakly associates with this fraction. Therefore, a calcium-myristoyl switch seems to be a major, but not sole determinant for the association of VILIP with membranes in living cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Calcium / pharmacology*
  • Calcium-Binding Proteins / metabolism*
  • Immunohistochemistry
  • Nerve Tissue Proteins / metabolism*
  • Neurocalcin
  • Neurons / drug effects*
  • Neurons / metabolism
  • PC12 Cells / metabolism*
  • Rats
  • Receptors, Calcium-Sensing*
  • Transfection

Substances

  • Calcium-Binding Proteins
  • Nerve Tissue Proteins
  • Neurocalcin
  • Receptors, Calcium-Sensing
  • Vsnl1 protein, rat
  • Calcium