title: Sequence, chromophore extraction and 3-D model of the photoactive yellow protein from Rhodobacter sphaeroides.
abstract: The photoactive yellow protein (pyp) gene has been isolated from Rhodobacter sphaeroides by probing with a homologous PCR-product. A sequence analysis shows that this pyp gene encodes a 124 AA protein with 48% identity to the three known PYPs. Downstream from pyp, a number of adjacent open reading frames were identified, including a gene encoding a CoA-ligase homologue (pCL). This latter protein is proposed to be involved in PYP chromophore activation, required for attachment to the apoprotein. We have demonstrated the presence of the chromophoric group, previously identified in PYP from Ectothiorhodospira halophila as trans 4-hydroxy cinnamic acid, in phototrophically cultured R. sphaeroides cells by capillary zone electrophoresis. The basic structure of the chromophore binding pocket in PYP has been conserved, as shown by a 3D model of R. sphaeroides PYP, constructed by homology-based molecular modelling. In addition, this model shows that R. sphaeroides PYP contains a characteristic, positively charged patch.
authors.affiliation: Laboratory for Microbiology, EC Slater Institute, University of Amsterdam, Nieuwe Achtergracht 127, 1018 WS Amsterdam, Netherlands.
authors.forename: R
authors.initials: R
authors.lastname: Kort
authors.forename: M K
authors.initials: MK
authors.lastname: Phillips-Jones
authors.forename: D M
authors.initials: DM
authors.lastname: van Aalten
authors.forename: A
authors.initials: A
authors.lastname: Haker
authors.forename: S M
authors.initials: SM
authors.lastname: Hoffer
authors.forename: K J
authors.initials: KJ
authors.lastname: Hellingwerf
authors.forename: W
authors.initials: W
authors.lastname: Crielaard
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D003373:Coumaric Acids / Q000737:chemistry*; D019075:Electrophoresis, Capillary; D008958:Models, Molecular; D008969:Molecular Sequence Data; D020283:Photoreceptors, Microbial; D016133:Polymerase Chain Reaction; D011422:Propionates; D011487:Protein Conformation; D012242:Rhodobacter sphaeroides / Q000378:metabolism*; D016415:Sequence Alignment
chemical_list: D001426:Bacterial Proteins; D003373:Coumaric Acids; D020283:Photoreceptors, Microbial; D011422:Propionates; C050162:photoactive yellow protein, Bacteria; C495469:p-coumaric acid