title: Secondary structure and folding of a functional chloroplast precursor protein.
abstract: Ferredoxin is a chloroplast stroma protein which is cytosolically synthesized as a precursor with an amino-terminal extension called the transit sequence that is needed for the post-translational uptake by the chloroplast. To characterize the secondary and tertiary structure elements, the full precursor, the holo- and apo- (without iron-sulfur cluster) forms of the mature protein, and the chemically synthesized transit peptide were obtained and analyzed separately. Circular dichroism, tryptophan fluorescence quenching, and protease accessibility experiments indicate that the precursor has a low content of defined secondary structure and resembles unfolded proteins; these properties are due to both the mature part and the transit sequence. This result provides an explanation for the lack of cytosolic factor requirement of this protein for import. In an import competition assay, the isolated transit peptide had an affinity for the chloroplasts comparable to the full precursor. Interestingly and of possible importance to the import process, the transit peptide has conformational flexibility as it adopts alternative secondary structures in different environments.
authors.affiliation: Institute of Molecular Biology and Medical Biotechnology, University of Utrecht, The Netherlands.
authors.forename: M
authors.initials: M
authors.lastname: Pilon
authors.forename: A G
authors.initials: AG
authors.lastname: Rietveld
authors.forename: P J
authors.initials: PJ
authors.lastname: Weisbeek
authors.forename: B
authors.initials: B
authors.lastname: de Kruijff
mesh_terms: D000595:Amino Acid Sequence; D001692:Biological Transport; D002736:Chloroplasts / Q000378:metabolism*; D002942:Circular Dichroism; D003600:Cytosol / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005288:Ferredoxins / Q000737:chemistry* / Q000378:metabolism; D008969:Molecular Sequence Data; D017510:Protein Folding; D011498:Protein Precursors / Q000737:chemistry* / Q000378:metabolism; D017433:Protein Structure, Secondary; D013050:Spectrometry, Fluorescence
chemical_list: D005288:Ferredoxins; D011498:Protein Precursors