title: Expression, isolation, and characterization of a chloroplast targeting peptide.
abstract: For the first time a method is described in which an N-terminal targeting peptide is isolated from Escherichia coli. After overexpression, purification, and cleavage of a fusion protein the protease-sensitive transit peptide from the chloroplast precursor protein preferredoxin could be isolated by HPLC. It was characterized by N-terminal amino acid sequencing and electrospray mass spectrometry. Its functionality was suggested by in vitro import competition experiments with isolated pea chloroplasts, in which the isolated peptide inhibited the import of radioactively labeled preferredoxin. Results from import competition experiments performed with a transit peptide deletion mutant suggested that the four extreme C-terminal amino acids lack information to interact with the chloroplast import machinery.
authors.affiliation: Centre for Biomembranes and Lipid Enzymology, Institute of Biomembranes, Utrecht University, Padualaan 8, Utrecht, 3584 CH, The Netherlands. h.l.j.wienk@chem.uu.nl
authors.forename: H L
authors.initials: HL
authors.lastname: Wienk
authors.forename: B
authors.initials: B
authors.lastname: de Kruijff
mesh_terms: D002736:Chloroplasts / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D004926:Escherichia coli / Q000378:metabolism; D005288:Ferredoxins / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification; D013058:Mass Spectrometry; D018532:Pisum sativum; D010940:Plant Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification; D011498:Protein Precursors / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification; D021382:Protein Sorting Signals / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification; D020539:Sequence Analysis, Protein
chemical_list: D005288:Ferredoxins; D010940:Plant Proteins; D011498:Protein Precursors; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins; C085169:preferredoxin