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Title: SET7/9 Catalytic Mutants Reveal the Role of Active Site Water Molecules in Lysine Multiple Methylation

Abstract

SET domain lysine methyltransferases (KMTs) methylate specific lysine residues in histone and non-histone substrates. These enzymes also display product specificity by catalyzing distinct degrees of methylation of the lysine {epsilon}-amino group. To elucidate the molecular mechanism underlying this specificity, we have characterized the Y245A and Y305F mutants of the human KMT SET7/9 (also known as KMT7) that alter its product specificity from a monomethyltransferase to a di- and a trimethyltransferase, respectively. Crystal structures of these mutants in complex with peptides bearing unmodified, mono-, di-, and trimethylated lysines illustrate the roles of active site water molecules in aligning the lysine {epsilon}-amino group for methyl transfer with S-adenosylmethionine. Displacement or dissociation of these solvent molecules enlarges the diameter of the active site, accommodating the increasing size of the methylated {epsilon}-amino group during successive methyl transfer reactions. Together, these results furnish new insights into the roles of active site water molecules in modulating lysine multiple methylation by SET domain KMTs and provide the first molecular snapshots of the mono-, di-, and trimethyl transfer reactions catalyzed by these enzymes.

Authors:
; ; ; ; ; ; ;  [1];  [2];  [2]
  1. (Michigan)
  2. (
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Org.:
USDOE
OSTI Identifier:
1002881
Resource Type:
Journal Article
Journal Name:
J. Biol. Chem.
Additional Journal Information:
Journal Volume: 285; Journal Issue: 10, 2010; Journal ID: ISSN 0021-9258
Country of Publication:
United States
Language:
ENGLISH
Subject:
36 MATERIALS SCIENCE; BEARINGS; CRYSTAL STRUCTURE; DISSOCIATION; ENZYMES; HISTONES; LYSINE; METHYLATION; MUTANTS; PEPTIDES; RESIDUES; SOLVENTS; SPECIFICITY; SUBSTRATES; TRANSFER REACTIONS; WATER

Citation Formats

Del Rizzo, Paul A., Couture, Jean-François, Dirk, Lynnette M.A., Strunk, Bethany S., Roiko, Marijo S., Brunzelle, Joseph S., Houtz, Robert L., Trievel, Raymond C., NWU), and Kentucky). SET7/9 Catalytic Mutants Reveal the Role of Active Site Water Molecules in Lysine Multiple Methylation. United States: N. p., 2010. Web. doi:10.1074/jbc.M110.114587.
Del Rizzo, Paul A., Couture, Jean-François, Dirk, Lynnette M.A., Strunk, Bethany S., Roiko, Marijo S., Brunzelle, Joseph S., Houtz, Robert L., Trievel, Raymond C., NWU), & Kentucky). SET7/9 Catalytic Mutants Reveal the Role of Active Site Water Molecules in Lysine Multiple Methylation. United States. doi:10.1074/jbc.M110.114587.
Del Rizzo, Paul A., Couture, Jean-François, Dirk, Lynnette M.A., Strunk, Bethany S., Roiko, Marijo S., Brunzelle, Joseph S., Houtz, Robert L., Trievel, Raymond C., NWU), and Kentucky). Mon . "SET7/9 Catalytic Mutants Reveal the Role of Active Site Water Molecules in Lysine Multiple Methylation". United States. doi:10.1074/jbc.M110.114587.
@article{osti_1002881,
title = {SET7/9 Catalytic Mutants Reveal the Role of Active Site Water Molecules in Lysine Multiple Methylation},
author = {Del Rizzo, Paul A. and Couture, Jean-François and Dirk, Lynnette M.A. and Strunk, Bethany S. and Roiko, Marijo S. and Brunzelle, Joseph S. and Houtz, Robert L. and Trievel, Raymond C. and NWU) and Kentucky)},
abstractNote = {SET domain lysine methyltransferases (KMTs) methylate specific lysine residues in histone and non-histone substrates. These enzymes also display product specificity by catalyzing distinct degrees of methylation of the lysine {epsilon}-amino group. To elucidate the molecular mechanism underlying this specificity, we have characterized the Y245A and Y305F mutants of the human KMT SET7/9 (also known as KMT7) that alter its product specificity from a monomethyltransferase to a di- and a trimethyltransferase, respectively. Crystal structures of these mutants in complex with peptides bearing unmodified, mono-, di-, and trimethylated lysines illustrate the roles of active site water molecules in aligning the lysine {epsilon}-amino group for methyl transfer with S-adenosylmethionine. Displacement or dissociation of these solvent molecules enlarges the diameter of the active site, accommodating the increasing size of the methylated {epsilon}-amino group during successive methyl transfer reactions. Together, these results furnish new insights into the roles of active site water molecules in modulating lysine multiple methylation by SET domain KMTs and provide the first molecular snapshots of the mono-, di-, and trimethyl transfer reactions catalyzed by these enzymes.},
doi = {10.1074/jbc.M110.114587},
journal = {J. Biol. Chem.},
issn = {0021-9258},
number = 10, 2010,
volume = 285,
place = {United States},
year = {2010},
month = {11}
}