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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.

SUBMITTER: Korennykh AV

PROVIDER: S-EPMC1847776 | biostudies-literature | 2006 May

REPOSITORIES: biostudies-literature

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The electrostatic character of the ribosomal surface enables extraordinarily rapid target location by ribotoxins.

Korennykh Alexei V AV Piccirilli Joseph A JA Correll Carl C CC

Nature structural & molecular biology 20060409 5

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 ...[more]

PMID: 16604082

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Project description:PurposeThe goal of this work is to produce a surface-dosimetry method capable of accurately and remotely measuring skin dose for patients undergoing total skin electron therapy (TSET) without the need for postexposure dosimeter processing. A rapid and wireless surface-dosimetry system was developed to improve clinical workflow. Scintillator-surface dosimetry was conducted on patients undergoing TSET by imaging scintillator targets with an intensified camera during TSET delivery.Methods and materialsDisc-shaped scintillator targets were attached to the skin surface of patients undergoing TSET and imaged with an intensified, time-gated, and linear accelerator-synchronized camera. Optically stimulated luminescence dosimeters (OSLDs) were placed directly adjacent to scintillators at several dosimetry sites to serve as an absolute dose reference. Real-time image-processing methods were used to produce background-subtracted intensity maps of Cherenkov and scintillation emission. Rapid conversion of scintillator-light output to dose was achieved by using a custom fitting algorithm and calibration factor. Surface doses measured by scintillators were compared with those from OSLDs.ResultsAbsolute surface-dose measurements for 99 dosimetry sites were evaluated. According to paired OSLD estimates, scintillator dosimeters were able to report dose with <3% difference in 88 of 99 observed dosimetry sites and <5% difference in 98 of 99 observed dosimetry sites. Fitting a linear regression to dose data reported by scintillator versus OSLD, per dosimetry site, yielded an R2 = 0.94.ConclusionsScintillators were able to report dose within <3% accuracy of OSLDs. Imaging of calibrated scintillator targets via an intensified, linear accelerator-synchronized camera provides rapid absolute surface-dosimetry measurements for patients treated with TSET. This technique has the potential to reduce the amount of time and effort necessary to conduct full-body dosimetry and can be adopted for use in any surface-dosimetry setting where the region of interest is observable throughout treatment.

Dataset Information

The electrostatic character of the ribosomal surface enables extraordinarily rapid target location by ribotoxins.

Publications

The electrostatic character of the ribosomal surface enables extraordinarily rapid target location by ribotoxins.

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