title: The electrostatic character of the ribosomal surface enables extraordinarily rapid target location by ribotoxins.
abstract: Alpha-sarcin ribotoxins comprise a unique family of ribonucleases that cripple the ribosome by catalyzing endoribonucleolytic cleavage of ribosomal RNA at a specific location in the sarcin/ricin loop (SRL). The SRL structure alone is cleaved site-specifically by the ribotoxin, but the ribosomal context enhances the reaction rate by several orders of magnitude. We show that, for the alpha-sarcin-like ribotoxin restrictocin, this catalytic advantage arises from favorable electrostatic interactions with the ribosome. Restrictocin binds at many sites on the ribosomal surface and under certain conditions cleaves the SRL with a second-order rate constant of 1.7 x 10(10) M(-1) s(-1), a value that matches the predicted frequency of random restrictocin-ribosome encounters. The results suggest a mechanism of target location whereby restrictocin encounters ribosomes randomly and diffuses within the ribosomal electrostatic field to the SRL. These studies show a role for electrostatics in protein-ribosome recognition.
authors.affiliation: Department of Chemistry, The University of Chicago, Chicago, Illinois 60637, USA.
authors.forename: Alexei V
authors.initials: AV
authors.lastname: Korennykh
authors.forename: Joseph A
authors.initials: JA
authors.lastname: Piccirilli
authors.forename: Carl C
authors.initials: CC
authors.lastname: Correll
mesh_terms: D000818:Animals; D001665:Binding Sites; D018360:Crystallography, X-Ray; D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D004722:Endoribonucleases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D005656:Fungal Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D008958:Models, Molecular; D009154:Mutation / Q000235:genetics; D009690:Nucleic Acid Conformation; D011487:Protein Conformation; D051381:Rats; D012270:Ribosomes / Q000737:chemistry* / Q000378:metabolism*; D055672:Static Electricity; D013379:Substrate Specificity; D013997:Time Factors; D017186:Titrimetry
chemical_list: D005656:Fungal Proteins; C014801:alpha-sarcin; D004247:DNA; D004722:Endoribonucleases