Design of peptide-acridine mimics of ribonuclease activity

Ching Hsuan Tung, Ziping Wei, Michael J Leibowitz, Stanley Stein

Research output: Contribution to journalArticle

61 Citations (Scopus)

Abstract

A series of peptide-acridine conjugates was designed and synthesized, based on three features of the proposed catalytic mechanism of RNase A: 2prime;-proton abstraction by His-12, proton donation to the leaving 5′-oxygen by His-119, and stabilization of the pentacoordinated phosphorous transition state by Lys-41. The substrate binding capability of RNase A was mimicked by the intercalator, acridine. Lysine served as a linker between acridine and the catalytic tripeptide. Cleavage of target RNA was monitored by agarose gel electrophoresis and by gel-permeation chromatography. The carboxyl-amidated conjugates HGHK(Acr)-NH2, HPHK(Acr)-NH2, and GGHK(Acr)-NH2 (where Acr indicates 2-methyl-9-acridinemethylene) all had similar hydrolytic activity. The catalytic mechanism most likely involved only the abstraction of the 2′-proton and stabilization of the transition state. These RNase mimics utilized rRNA and single-stranded RNA but not double-stranded RNA and tRNA as substrates.

Original languageEnglish (US)
Pages (from-to)7114-7118
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume89
Issue number15
StatePublished - 1992
Externally publishedYes

Fingerprint

Acridines
Ribonucleases
Protons
Pancreatic Ribonuclease
Peptides
RNA Cleavage
Intercalating Agents
Double-Stranded RNA
Agar Gel Electrophoresis
Transfer RNA
Lysine
Gel Chromatography
Oxygen

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Design of peptide-acridine mimics of ribonuclease activity. / Tung, Ching Hsuan; Wei, Ziping; Leibowitz, Michael J; Stein, Stanley.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 89, No. 15, 1992, p. 7114-7118.

Research output: Contribution to journalArticle

@article{682736a74fe8497c840823fa702920d6,
title = "Design of peptide-acridine mimics of ribonuclease activity",
abstract = "A series of peptide-acridine conjugates was designed and synthesized, based on three features of the proposed catalytic mechanism of RNase A: 2prime;-proton abstraction by His-12, proton donation to the leaving 5′-oxygen by His-119, and stabilization of the pentacoordinated phosphorous transition state by Lys-41. The substrate binding capability of RNase A was mimicked by the intercalator, acridine. Lysine served as a linker between acridine and the catalytic tripeptide. Cleavage of target RNA was monitored by agarose gel electrophoresis and by gel-permeation chromatography. The carboxyl-amidated conjugates HGHK(Acr)-NH2, HPHK(Acr)-NH2, and GGHK(Acr)-NH2 (where Acr indicates 2-methyl-9-acridinemethylene) all had similar hydrolytic activity. The catalytic mechanism most likely involved only the abstraction of the 2′-proton and stabilization of the transition state. These RNase mimics utilized rRNA and single-stranded RNA but not double-stranded RNA and tRNA as substrates.",
author = "Tung, {Ching Hsuan} and Ziping Wei and Leibowitz, {Michael J} and Stanley Stein",
year = "1992",
language = "English (US)",
volume = "89",
pages = "7114--7118",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
number = "15",

}

TY - JOUR

T1 - Design of peptide-acridine mimics of ribonuclease activity

AU - Tung, Ching Hsuan

AU - Wei, Ziping

AU - Leibowitz, Michael J

AU - Stein, Stanley

PY - 1992

Y1 - 1992

N2 - A series of peptide-acridine conjugates was designed and synthesized, based on three features of the proposed catalytic mechanism of RNase A: 2prime;-proton abstraction by His-12, proton donation to the leaving 5′-oxygen by His-119, and stabilization of the pentacoordinated phosphorous transition state by Lys-41. The substrate binding capability of RNase A was mimicked by the intercalator, acridine. Lysine served as a linker between acridine and the catalytic tripeptide. Cleavage of target RNA was monitored by agarose gel electrophoresis and by gel-permeation chromatography. The carboxyl-amidated conjugates HGHK(Acr)-NH2, HPHK(Acr)-NH2, and GGHK(Acr)-NH2 (where Acr indicates 2-methyl-9-acridinemethylene) all had similar hydrolytic activity. The catalytic mechanism most likely involved only the abstraction of the 2′-proton and stabilization of the transition state. These RNase mimics utilized rRNA and single-stranded RNA but not double-stranded RNA and tRNA as substrates.

AB - A series of peptide-acridine conjugates was designed and synthesized, based on three features of the proposed catalytic mechanism of RNase A: 2prime;-proton abstraction by His-12, proton donation to the leaving 5′-oxygen by His-119, and stabilization of the pentacoordinated phosphorous transition state by Lys-41. The substrate binding capability of RNase A was mimicked by the intercalator, acridine. Lysine served as a linker between acridine and the catalytic tripeptide. Cleavage of target RNA was monitored by agarose gel electrophoresis and by gel-permeation chromatography. The carboxyl-amidated conjugates HGHK(Acr)-NH2, HPHK(Acr)-NH2, and GGHK(Acr)-NH2 (where Acr indicates 2-methyl-9-acridinemethylene) all had similar hydrolytic activity. The catalytic mechanism most likely involved only the abstraction of the 2′-proton and stabilization of the transition state. These RNase mimics utilized rRNA and single-stranded RNA but not double-stranded RNA and tRNA as substrates.

UR - http://www.scopus.com/inward/record.url?scp=0026780554&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0026780554&partnerID=8YFLogxK

M3 - Article

C2 - 1379732

AN - SCOPUS:0026780554

VL - 89

SP - 7114

EP - 7118

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 15

ER -