Active-site alkylation destabilizes human O6-alkylguanine DNA alkyltransferase

Joseph J. Rasimas; Paula A. Dalessio; Ira J. Ropson; Anthony E. Pegg; Michael G. Fried

doi:10.1110/ps.03319404

Active-site alkylation destabilizes human O⁶-alkylguanine DNA alkyltransferase

Joseph J. Rasimas, Paula A. Dalessio, Ira J. Ropson, Anthony E. Pegg, Michael G. Fried

Research output: Contribution to journal › Article › peer-review

26 Scopus citations

Abstract

O⁶-alkylguanine-DNA alkyltransferase (AGT) repairs pro-mutagenic O⁶-alkylguanine and O⁴-alkylthymine lesions in DNA. The alkylated form of the protein is not reactivated; instead, it is rapidly ubiquitinated and degraded. Here, we show that alkylation destabilizes the native fold of the protein by 0.5-1.2 kcal/mole and the DNA-binding function by 0.8-1.4 kcal/mole. On this basis, we propose that destabilization of the native conformational ensemble acts as a signal for ubiquitination.

Original language	English
Pages (from-to)	301-305
Number of pages	5
Journal	Protein Science
Volume	13
Issue number	1
DOIs	https://doi.org/10.1110/ps.03319404
State	Published - Jan 2004

Keywords

DNA binding
Denaturation
O -methylguanine-methyltransferase
O-alkylguanine-DNA alkyltransferase
Protein-alkylation

ASJC Scopus subject areas

Biochemistry
Molecular Biology

Access to Document

10.1110/ps.03319404

Cite this

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title = "Active-site alkylation destabilizes human O6-alkylguanine DNA alkyltransferase",

abstract = "O6-alkylguanine-DNA alkyltransferase (AGT) repairs pro-mutagenic O6-alkylguanine and O4-alkylthymine lesions in DNA. The alkylated form of the protein is not reactivated; instead, it is rapidly ubiquitinated and degraded. Here, we show that alkylation destabilizes the native fold of the protein by 0.5-1.2 kcal/mole and the DNA-binding function by 0.8-1.4 kcal/mole. On this basis, we propose that destabilization of the native conformational ensemble acts as a signal for ubiquitination.",

keywords = "DNA binding, Denaturation, O -methylguanine-methyltransferase, O-alkylguanine-DNA alkyltransferase, Protein-alkylation",

author = "Rasimas, {Joseph J.} and Dalessio, {Paula A.} and Ropson, {Ira J.} and Pegg, {Anthony E.} and Fried, {Michael G.}",

year = "2004",

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language = "English",

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T1 - Active-site alkylation destabilizes human O6-alkylguanine DNA alkyltransferase

AU - Rasimas, Joseph J.

AU - Dalessio, Paula A.

AU - Ropson, Ira J.

AU - Pegg, Anthony E.

AU - Fried, Michael G.

PY - 2004/1

Y1 - 2004/1

N2 - O6-alkylguanine-DNA alkyltransferase (AGT) repairs pro-mutagenic O6-alkylguanine and O4-alkylthymine lesions in DNA. The alkylated form of the protein is not reactivated; instead, it is rapidly ubiquitinated and degraded. Here, we show that alkylation destabilizes the native fold of the protein by 0.5-1.2 kcal/mole and the DNA-binding function by 0.8-1.4 kcal/mole. On this basis, we propose that destabilization of the native conformational ensemble acts as a signal for ubiquitination.

AB - O6-alkylguanine-DNA alkyltransferase (AGT) repairs pro-mutagenic O6-alkylguanine and O4-alkylthymine lesions in DNA. The alkylated form of the protein is not reactivated; instead, it is rapidly ubiquitinated and degraded. Here, we show that alkylation destabilizes the native fold of the protein by 0.5-1.2 kcal/mole and the DNA-binding function by 0.8-1.4 kcal/mole. On this basis, we propose that destabilization of the native conformational ensemble acts as a signal for ubiquitination.

KW - DNA binding

KW - Denaturation

KW - O -methylguanine-methyltransferase

KW - O-alkylguanine-DNA alkyltransferase

KW - Protein-alkylation

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