title: Secondary structures of a new class of lipid body proteins from oilseeds.
abstract: The three main isoforms of the 19-kDa lipid body proteins (oleosin) have been purified to homogeneity from embryos of rapeseed. The secondary structures of these proteins as derived from circular dichroism (CD) and Fourier transform infrared (FTIR) spectroscopy were compared with the secondary structures predicted from the primary sequences. The salient feature of the primary sequence of all oleosins is its division into three defined structural domains: a central hydrophobic domain flanked on either side by relatively hydrophilic domains, respectively. Using a variety of predictive methods based on primary amino acid sequence data, the oleosins exhibited a high probability of beta-strand structure in the 70-residue central hydrophobic domain, with relatively little alpha-helical content. Secondary structure data derived from CD and FTIR were consistent with the predictions from primary sequence, showing that the oleosins contained about 45% beta-strand and 13% alpha-helical structure. Under high salt conditions, a 40-kDa polypeptide was obtained from purified preparations of the 19-kDa oleosins. The 40-kDa polypeptide has a very similar secondary structure, as analyzed by CD and FTIR, to that of the 19-kDa oleosins. This polypeptide is therefore probably a dimer of the 19-kDa oleosins that is formed in high salt environments. A model of the general structure of oleosins is proposed whereby the central hydrophobic domain of the protein with a predominantly beta-strand structure is embedded into the non-aqueous phase of lipid-bodies. This hydrophobic region is flanked by putative alpha-helical structures in the polar N- and C-terminal domains which are probably oriented at the lipid-water interface.
authors.affiliation: Department of Brassica and Oilseeds Research, Cambridge Laboratory, John Innes Center for Plant Science Research, Norwich, United Kingdom.
authors.forename: M
authors.initials: M
authors.lastname: Li
authors.forename: L J
authors.initials: LJ
authors.lastname: Smith
authors.forename: D C
authors.initials: DC
authors.lastname: Clark
authors.forename: R
authors.initials: R
authors.lastname: Wilson
authors.forename: D J
authors.initials: DJ
authors.lastname: Murphy
mesh_terms: D000595:Amino Acid Sequence; D001937:Brassica / Q000378:metabolism*; D002850:Chromatography, Gel; D002942:Circular Dichroism; D004591:Electrophoresis, Polyacrylamide Gel; D005583:Fourier Analysis; D008074:Lipoproteins / Q000235:genetics* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000235:genetics* / Q000302:isolation & purification; D011487:Protein Conformation; D013055:Spectrophotometry, Infrared; D013056:Spectrophotometry, Ultraviolet
chemical_list: D008074:Lipoproteins; D010940:Plant Proteins