title: 1H-NMR and photo-CIDNP spectroscopies show a possible role for Trp23 and Phe31 in nucleic acid binding by P2 ribonuclease from the archaeon Sulfolobus solfataricus.
abstract: Investigations were performed on recombinant ribonuclease P2 from Sulfolobus solfataricus, previously cloned and expressed in Escherichia coli [Fusi, P., Grisa, M., Mombelli, E., Consonni, R., Tortora, P. and Vanoni, M. (1995) Gene 154, 99-103]. NMR and photo-CIDNP spectroscopies showed that the enzyme possesses an aromatic cluster consisting of Phe5, Tyr7, Phe31 and Tyr33 while Trp23 is fully exposed to solvent. Phe31, Tyr33 and Trp23 are located within a triple stranded antiparallel beta-sheet, each one being part of an amino acid stretch matching consensus sequences for RNA binding. Phe31 and Trp23 are exposed to and specifically interact with a flavin dye used as a model ligand, with a topology reminiscent of that found in several eubacterial and eukariotic RNA-binding proteins.
authors.affiliation: Istituto di Chimica delle Macromolecole, Lab. NMR, CNR Milano, Italy.
authors.forename: R
authors.initials: R
authors.lastname: Consonni
authors.forename: R
authors.initials: R
authors.lastname: Limiroli
authors.forename: H
authors.initials: H
authors.lastname: Molinari
authors.forename: P
authors.initials: P
authors.lastname: Fusi
authors.forename: M
authors.initials: M
authors.lastname: Grisa
authors.forename: M
authors.initials: M
authors.lastname: Vanoni
authors.forename: P
authors.initials: P
authors.lastname: Tortora
mesh_terms: D001665:Binding Sites; D004926:Escherichia coli / Q000235:genetics; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D010649:Phenylalanine; D012313:RNA / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012260:Ribonucleases / Q000235:genetics / Q000378:metabolism*; D013057:Spectrum Analysis / Q000379:methods; D016944:Sulfolobus / Q000201:enzymology*; D014364:Tryptophan
chemical_list: D011994:Recombinant Proteins; D010649:Phenylalanine; D012313:RNA; D014364:Tryptophan; D012260:Ribonucleases; C092231:ribonuclease P2