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1 H‐NMR and photo‐CIDNP spectroscopies show a possible role for Trp 23 and Phe 31 in nucleic acid binding by P2 ribonuclease from the archaeon Sulfolobus solfataricus
Author(s) -
Consonni Roberto,
Limiroli Rita,
Molinari Henriette,
Fusi Paola,
Grisa Margareth,
Vai Marco,
Tortora Paolo
Publication year - 1995
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(95)00940-b
Subject(s) - cidnp , ribonuclease , nucleic acid , chemistry , nuclear magnetic resonance spectroscopy , rna , biochemistry , stereochemistry , polarization (electrochemistry) , gene
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 constisting of Phe 5 , Tyr 7 , Phe 31 and Tyr 33 while Trp 23 is fully exposed to solvent. Phe 31 , Tyr 33 and Trp 23 are located within a triple stranded antiparallel β‐sheet, each one being part of an amino acid stretch matching consensus sequences for RNA binding. Phe 31 and Trp 23 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.