title: Reconstitution of mature plastocyanin from precursor apo-plastocyanin expressed in Escherichia coli.
abstract: The precursor plastocyanin from Silene pratensis (white campion) has been expressed in Escherichia coli. The precursor protein was accumulated in insoluble aggregates and partially purified as an apo-protein. The purified precursor apo-plastocyanin was processed to the mature apo-plastocyanin by chloroplast extracts. N-terminal amino-acid sequencing indicated that the processed protein was identical to the N-terminal amino-acid residues of mature plastocyanin that was deduced from the nucleotide sequence. The copper could be incorporated into the apo-plastocyanin of mature size in vitro, but could not into the precursor apo-plastocyanin under the same conditions. Absorption spectra and reduction potential of the reconstituted mature plastocyanin were indistinguishable from those of the purified spinach plastocyanin. The electron transfer activities of the reconstituted plastocyanin with both the Photosystem I reaction center (P700) and cytochrome f were almost the same as those of the purified spinach plastocyanin.
authors.affiliation: Department of Chemistry, Faculty of Science and Technology, Meijo University, Nagoya, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Hibino
authors.forename: A
authors.initials: A
authors.lastname: Douwe de Boer
authors.forename: P J
authors.initials: PJ
authors.lastname: Weisbeek
authors.forename: T
authors.initials: T
authors.lastname: Takabe
mesh_terms: D000595:Amino Acid Sequence; D001059:Apoproteins / Q000378:metabolism*; D015153:Blotting, Western; D003300:Copper / Q000378:metabolism; D003580:Cytochromes / Q000737:chemistry; D045348:Cytochromes f; D004579:Electron Transport; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D007700:Kinetics; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D010777:Photochemistry; D010944:Plants / Q000378:metabolism; D010957:Plasmids; D010970:Plastocyanin / Q000302:isolation & purification / Q000378:metabolism*; D011498:Protein Precursors / Q000378:metabolism*
chemical_list: D001059:Apoproteins; D003580:Cytochromes; D011498:Protein Precursors; C040778:apoplastocyanin; D003300:Copper; D010970:Plastocyanin; D045348:Cytochromes f