Identification of proteins associated with plastoglobules isolated from pea (Pisum sativum L.) chloroplasts.
Plastoglobules are conspicuous lipid-containing structures in the chloroplast stroma and are thought to serve as lipid reservoirs for thylakoid membranes. Plastoglobules also contain low levels of proteins. We have purified plastoglobules from a pea chloroplast membrane fraction. Sodium dodecyl sulfate-polyacrylamide gel electrophoretic analysis of the purified plastoglobules revealed more than a dozen distinct polypeptides that we propose to term plastoglobulins. For one of the proteins, termed plastoglobulin 1 (PG1), we have obtained partial N-terminal and internal protein sequences. The amino acid sequence, deduced from cDNA, encoded a precursor protein of a calculated mass of 38,491 Da which contained a 48-residue-long N-terminal signal sequence. Pre-PG1 obtained by coupled in-vitro transcription/translation was imported into pea chloroplasts, its signal sequence was cleaved and mature PG1 was assembled into plastoglobules. Homology searches of the data bases revealed similarity of PG1 to the plastoglobule-associated protein of Capsicum annuum, the carotenoid-associated protein of Cucumis sativus and to a protein of the cyanobacterium Synechocystis sp., indicating that PG1 is a novel member of an ancient protein family. Immunoelectron microscopy using PG1 specific antibodies indicated that PG1 is present in multiple copies on the surface of plastoglobules.
Kessler
F
F
Institute of Plant Sciences, Swiss Federal Institute of Technology (ETH), Zürich, Switzerland. felix.kessler@ipw.biol.ethz.ch
Schnell
D
D
Blobel
G
G
D000595:Amino Acid Sequence; D001483:Base Sequence; D002736:Chloroplasts / Q000378:metabolism* / Q000648:ultrastructure; D003001:Cloning, Molecular; D017931:DNA Primers; D016253:Microscopy, Immunoelectron / Q000379:methods; D008969:Molecular Sequence Data; D015388:Organelles / Q000378:metabolism*; D018532:Pisum sativum / Q000378:metabolism; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid
D017931:DNA Primers; C118749:PG1 protein, Pisum sativum; D010940:Plant Proteins