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Journal article
Enhancing ubiquitin crystallization through surfaceentropy reduction
Acta crystallographica. Section F, Structural biology communications, v 70, pp 1434-1442
01 Oct 2014
PMID: 25286958
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
Ubiquitin has many attributes suitable for a crystallization chaperone, including high stability and ease of expression. However, ubiquitin contains a high surface density of lysine residues and the doctrine of surface-entropy reduction suggests that these lysines will resist participating in packing interactions and thereby impede crystallization. To assess the contributions of these residues to crystallization behavior, each of the seven lysines of ubiquitin was mutated to serine and the corresponding single-site mutant proteins were expressed and purified. The behavior of these seven mutants was then compared with that of the wild-type protein in a 384-condition crystallization screen. The likelihood of obtaining crystals varied by two orders of magnitude within this set of eight proteins. Some mutants crystallized much more readily than the wild type, while others crystallized less readily. X-ray crystal structures were determined for three readily crystallized variants: K11S, K33S and the K11S/K63S double mutant. These structures revealed that the mutant serine residues can directly promote crystallization by participating in favorable packing interactions; the mutations can also exert permissive effects, wherein crystallization appears to be driven by removal of the lysine rather than by addition of a serine. Presumably, such permissive effects reflect the elimination of steric and electrostatic barriers to crystallization.
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Details
- Title
- Enhancing ubiquitin crystallization through surfaceentropy reduction
- Creators
- Patrick J. Loll - Drexel UniversityPeining Xu - Drexel UniversityJohn T. Schmidt - Drexel UniversityScott L. Melideo - Drexel University
- Publication Details
- Acta crystallographica. Section F, Structural biology communications, v 70, pp 1434-1442
- Publisher
- Int Union Crystallography
- Number of pages
- 9
- Grant note
- Department of Biochemistry and Molecular Biology at the Drexel University College of Medicine R01GM079508 / NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Institute of General Medical Sciences (NIGMS) R01GM079508 / NIH/NIGMS; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Institute of General Medical Sciences (NIGMS) DE-AC02-06CH11357 / US Department of Energy, Office of Science, Office of Basic Energy Sciences; United States Department of Energy (DOE)
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Biochemistry and Molecular Biology
- Web of Science ID
- WOS:000343060200029
- Scopus ID
- 2-s2.0-84927563994
- Other Identifier
- 991019170584904721
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InCites Highlights
Data related to this publication, from InCites Benchmarking & Analytics tool:
- Web of Science research areas
- Biochemical Research Methods
- Biochemistry & Molecular Biology
- Biophysics
- Crystallography