Import of a new chloroplast inner envelope protein is greatly stimulated by potassium phosphate.
A cDNA clone encoding a major chloroplast inner envelope membrane protein of 96 kDa (IEP96) was isolated and characterized. The protein is synthesized as a larger-molecular-weight precursor (pIEP96) which contains a cleavable N-terminal transit sequence of 50 amino acids. The transit peptide exhibits typical stromal targeting information. It is cleaved in vitro by the stromal processing peptidase, though the mature protein is clearly localized in the inner envelope membrane. Translocation of pIEP96 into chloroplasts is greatly stimulated in the presence of 80 mM potassium phosphate which results in an import efficiency of about 90%. This effect is specific for potassium and phosphate, but cannot be ascribed to a membrane potential across the inner envelope membrane. Protein sequence analysis reveals five stretches of repeats of 26 amino acids in length. The N-terminal 300 amino acids are 45% identical (76% similarity) to the 35 kDa alpha-subunit of acetyl-CoA carboxyl-transferase from Escherichia coli. The C-terminal 500 amino acids share significant similarity (69%) with USOI, a component of the cytoskeleton in yeast.
Hirsch
S
S
Botanisches Institut, Universität Kiel, Germany.
Soll
J
J
D000595:Amino Acid Sequence; D001692:Biological Transport / Q000187:drug effects; D060365:Chloroplast Proteins; D002736:Chloroplasts / Q000378:metabolism*; D003001:Cloning, Molecular; D007425:Intracellular Membranes / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D018532:Pisum sativum / Q000235:genetics / Q000378:metabolism; D010710:Phosphates / Q000494:pharmacology*; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D017680:Potassium Compounds / Q000494:pharmacology*
D060365:Chloroplast Proteins; D008565:Membrane Proteins; D010710:Phosphates; D010940:Plant Proteins; D017680:Potassium Compounds; C013216:potassium phosphate