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Import of a new chloroplast inner envelope protein is greatly stimulated by potassium phosphate

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Abstract

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 α-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.

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Abbreviations

Pi :

phosphate

IEP:

inner envelope membrane protein

pIEP:

precursor form of IEP

SSU:

small subunit of ribulose-1,5-bisphosphate carboxylase oxygenase

αIEP96pep :

peptide specific antiserum to IEP96

αIEP96pol :

polyspecific antiserum to IEP96

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  1. Botanisches Institut, Universität Kiel, 24098, Kiel, Germany

    Stephan Hirsch & Jürgen Soll

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  1. Stephan Hirsch
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  2. Jürgen Soll
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Hirsch, S., Soll, J. Import of a new chloroplast inner envelope protein is greatly stimulated by potassium phosphate. Plant Mol Biol 27, 1173–1181 (1995). https://doi.org/10.1007/BF00020890

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  • DOI: https://doi.org/10.1007/BF00020890

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