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Design of an expression system to enhance MBP-mediated crystallization

Journal Article · · Scientific Reports
DOI:https://doi.org/10.1038/srep40991· OSTI ID:1357647
 [1];  [2];  [2];  [3];  [3];  [3];  [2];  [2];  [4]
  1. Univ. of Science and Technology of China, Hefei (China); National Inst. of Health (NIH), Bethesda, MD (United States)
  2. National Inst. of Health (NIH), Bethesda, MD (United States)
  3. Univ. of Science and Technology of China, Hefei (China)
  4. Case Western Reserve Univ., Cleveland, OH (United States)
+ Show Author Affiliations

Crystallization chaperones have been used to facilitate the crystallization of challenging proteins. Even though the maltose-binding protein (MBP) is one of the most commonly used crystallization chaperones, the design of optimal expression constructs for crystallization of MBP fusion proteins remains a challenge. To increase the success rate of MBP-facilitated crystallization, a series of expression vectors have been designed with either a short flexible linker or a set of rigid helical linkers. Seven death domain superfamily members were tested for crystallization with this set of vectors, six of which had never been crystallized before. All of the seven targets were crystallized, and their structures were determined using at least one of the vectors. Our successful crystallization of all of the targets demonstrates the validity of our approach and expands the arsenal of the crystallization chaperone toolkit, which may be applicable to crystallization of other difficult protein targets, as well as to other crystallization chaperones.

Research Organization:
Argonne National Laboratory (ANL), Argonne, IL (United States)
Sponsoring Organization:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22); National Cancer Inst.; National Inst. of General Medical Sciences; China Postdoctoral Science Foundation
Grant/Contract Number:
AC02-98CH10886
OSTI ID:
1357647
Alternate ID(s):
OSTI ID: 1409538
Journal Information:
Scientific Reports, Journal Name: Scientific Reports Journal Issue: 1 Vol. 7; ISSN 2045-2322
Publisher:
Nature Publishing GroupCopyright Statement
Country of Publication:
United States
Language:
ENGLISH

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Cited By (12)

Passenger sequences can promote interlaced dimers in a common variant of the maltose-binding protein journal December 2019
Polyionic Tags as Enhancers of Protein Solubility in Recombinant Protein Expression journal May 2018
Evaluation of multiple fused partners on enhancing soluble level of prenyltransferase NovQ in Escherichia coli journal November 2018
Crystal structure of the Streptococcus agalactiae CAMP factor provides insights into its membrane-permeabilizing activity journal June 2018
T4 lysozyme-facilitated crystallization of the human molybdenum cofactor-dependent enzyme mARC journal May 2018
Crystal structures of the recombinant β-factor XIIa protease with bound Thr-Arg and Pro-Arg substrate mimetics journal June 2019
Crystal structures of the recombinant β-factor XIIa protease with bound Thr-Arg and Pro-Arg substrate mimetics text January 2019
Structure determination of the CAMP factor of Streptococcus agalactiae with the aid of an MBP tag and insights into membrane-surface attachment journal July 2019
Structure determination of the CAMP factor of Streptococcus agalactiae with the aid of an MBP tag and insights into membrane-surface attachment text January 2019
Functional and structural characterization of zebrafish ASC journal June 2018
Crystal structure and activation mechanism of DR3 death domain journal April 2019
T4 lysozyme-facilitated crystallization of the human molybdenum cofactor-dependent enzyme mARC text January 2018

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Related Subjects

59 BASIC BIOLOGICAL SCIENCES
X-ray crystallography