Import inhibition of poly(His) containing chloroplast precursor proteins by Ni2+ ions

FEBS Lett. 1997 Feb 10;403(1):15-8. doi: 10.1016/s0014-5793(97)00016-1.

Abstract

The precursor of the small subunit of ribulose-1,5-bisphosphate carboxylase (pSS) and a modified pSS containing a C-terminal hexahistidyl tail (pSS(His)6) were imported into isolated Chlamydomonas chloroplasts with comparable efficiency. In the presence of Ni2+ ions the import of pSS(His)6 was inhibited and the precursor bound to the envelope remained protease sensitive, while import of pSS was not affected. Addition of an excess of L-histidine suppressed the inhibition demonstrating that the hexahistidyl-Ni2+ complex was responsible for import inhibition. Inhibition could be observed between about 0.5 and 10 mM Ni2+, depending on the total protein content in the assay. Import incompetent Ni2+ -precursor complexes can be used to study early events in chloroplast protein import.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biological Transport / drug effects
  • Chlamydomonas / drug effects
  • Chlamydomonas / metabolism
  • Cloning, Molecular
  • Dose-Response Relationship, Drug
  • Histidine / chemistry
  • Histidine / genetics
  • Nickel / pharmacology*
  • Protein Precursors / drug effects
  • Protein Precursors / genetics
  • Protein Precursors / metabolism*
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Ribulose-Bisphosphate Carboxylase / drug effects
  • Ribulose-Bisphosphate Carboxylase / genetics*
  • Ribulose-Bisphosphate Carboxylase / metabolism*

Substances

  • Protein Precursors
  • Recombinant Fusion Proteins
  • Histidine
  • Nickel
  • Ribulose-Bisphosphate Carboxylase