title: Nonorganellar acyl carrier protein from oleaginous yeast is a homologue of ribosomal protein P2.
abstract: Acyl carrier protein (ACP) is responsible for carrying the growing fatty acid chain from one enzyme active site to the next during fatty acid biosynthesis. Here we report the identification, purification, immunocytochemical localization, and cloning of ACP from the oleaginous yeast, Rhodotorula glutinis. The soluble fraction of this organism can synthesize triacylglycerol and is able to accept the acyl group from acyl-ACP for the synthesis. The ACP, cloned from the system, showed a significant similarity with ribosomal protein P2. Expression and characterization of the recombinant protein showed that the ACP was acylated in vitro. The recombinant protein was post-translationally modified, since it was observed in [14C]beta-alanine labeling and matrix-assisted laser desorption mass spectroscopic analysis. Site-directed mutants were generated to identify a serine residue responsible for phosphopantetheinylation and found that mutation of serine 59 to alanine abrogated the fatty acylation ability of the protein. These results demonstrate that a novel modification of ribosomal protein P2 allows it to act as an acyl carrier protein and participate in acylation reactions.
authors.affiliation: Department of Biochemistry, Indian Institute of Science, Bangalore 560012, India.
authors.forename: Sumana
authors.initials: S
authors.lastname: Raychaudhuri
authors.forename: Ram
authors.initials: R
authors.lastname: Rajasekharan
mesh_terms: D000213:Acyl Carrier Protein / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D007150:Immunohistochemistry; D008969:Molecular Sequence Data; D010750:Phosphoproteins / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000302:isolation & purification; D012248:Rhodotorula / Q000737:chemistry*; D012269:Ribosomal Proteins; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D000213:Acyl Carrier Protein; D010750:Phosphoproteins; D011994:Recombinant Proteins; D012269:Ribosomal Proteins; C031937:phosphoprotein P2, ribosomal