Journal article

Publication year: 2004

Pages: 49338-49345

Journal: Journal of Biological Chemistry

Volume number: 279

Issue number: 47

ISSN: 0021-9258

Open access status: Hybrid

DOI Link: https://doi.org/10.1074/jbc.M409307200

Publisher: Elsevier

Abstract: 
Strand discrimination in Escherichia coli DNA mismatch repair requires the activation of the endonuclease MutH by MutL. There is evidence that MutH binds to the N-terminal domain of MutL in an ATP-dependent manner; however, the interaction sites and the molecular mechanism of MutH activation have not yet been determined. We used a combination of site-directed mutagenesis and site-specific cross-linking to identify protein interaction sites between the proteins MutH and MutL. Unique cysteine residues were introduced in cysteine-free variants of MutH and MutL. The introduced cysteines were modified with the cross-linking reagent 4-maleimidobenzophenone. Photoactivation resulted in cross-links verified by mass spectrometry of some of the single cysteine variants to their respective Cys-free partner proteins. Moreover, we mapped the site of interaction by cross-linking different combinations of single cysteine MutH and MutL variants with thiol-specific homobifunctional cross-linkers of varying length. These results were used to model the MutH.MutL complex and to explain the ATP dependence of this interaction.

Harvard Citation style: Giron-Monzon, L., Manelyte, L., Ahrends, R., Kirsch, D., Spengler, B. and Friedhoff, P. (2004) Mapping protein-protein interactions between MutL and MutH by cross-linking, Journal of Biological Chemistry, 279(47), pp. 49338-49345. https://doi.org/10.1074/jbc.M409307200

APA Citation style: Giron-Monzon, L., Manelyte, L., Ahrends, R., Kirsch, D., Spengler, B., & Friedhoff, P. (2004). Mapping protein-protein interactions between MutL and MutH by cross-linking. Journal of Biological Chemistry. 279(47), 49338-49345. https://doi.org/10.1074/jbc.M409307200