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PMID: 17098187 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Lysine methylation as a routine rescue strategy for protein crystallization.

Structure (London, England : 1993) ·Vol. 14 ·No. 11 ·2006-11-00 ·Pages 1617-22

Walter TS, Meier C, Assenberg R, Au KF, Ren J, Verma A, Nettleship JE, Owens RJ, Stuart DI, Grimes JM

Abstract

Crystallization remains a critical step in X-ray structure determination. Because it is not generally possible to rationally predict crystallization conditions, commercial screens have been developed which sample a wide range of crystallization space. While this approach has proved successful in many cases, a significant number of proteins fail to crystallize despite being soluble and monodispersed. It is established that chemical modification can facilitate the crystallization of otherwise intractable proteins. Here we describe a method for the reductive methylation of lysine residues which is simple, inexpensive, and efficient, and report on its application to ten proteins. We describe the effect of methylation on the physico-chemical properties of these proteins, and show that it led to diffraction-quality crystals from four proteins and structures for three that had hitherto proved refractory to crystallization. The method is suited to both low- and high-throughput laboratories.

MeSH Terms
Bacillus anthracis/enzymology Biochemistry/methods Cohort Studies Computational Biology Crystallization/instrumentation,methods Crystallography, X-Ray/instrumentation,methods Flavivirus/metabolism Humans Lysine/chemistry Methylation Models, Molecular Murine hepatitis virus/metabolism Protein Conformation Proteins/chemistry
Chemicals
Proteins Lysine
Authors & Affiliations
10 authors, click to expand affiliations / ORCID
Walter Thomas S
Oxford Protein Production Facility, The Henry Wellcome Building for Genomic Medicine, Oxford University, Roosevelt Drive, Oxford OX3 7BN, United Kingdom.
Meier Christoph
Assenberg Rene
Au Kin-Fai
Ren Jingshan
Verma Anil
Nettleship Joanne E
Owens Raymond J
Stuart David I
Grimes Jonathan M
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Article Info
Journal
Structure (London, England : 1993)
Abbr.
Structure
ISSN
0969-2126
Published
2006-11-00
Pages
1617-22
Language
English
Region
United States
NLM ID
101087697
PMCID
PMC7126202
Subset
IM
Grants
Wellcome Trust · United Kingdom
Medical Research Council · G0400717 · United Kingdom
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