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Title: A Single Point Mutation Changes in the Crystallization Behavior of Mycoplasma arthritidis-derived Mitogen

Abstract

Mycoplasma arthritidis-derived mitogen (MAM) functions as a conventional superantigen (SAg). Although recombinant MAM has been crystallized by the hanging-drop vapor-diffusion method, the crystals diffracted poorly to only 5.0 Angstroms resolution, with large unit-cell parameters a = 163.8, b = 93.0, c = 210.9 Angstroms, {beta} = 93.7 degrees in the monoclinic space group P2{sub 1}. Unit-cell content analysis revealed that as many as 24 molecules could be present in the asymmetric unit. Systematic alanine mutagenesis was applied in order to search for mutants that give crystals of better quality. Two mutants, L50A and K201A, were crystallized under the same conditions as wild-type MAM (MAM{sub wt}). Crystals of the L50A mutant are isomorphous with those of MAMwt, while a new crystal form was obtained for the K201 mutant, belonging to the cubic space group P4132 with unit-cell parameters a = b = c = 181.9 Angstroms. Diffraction data were collected to 3.6 and 2.8 Angstroms resolution from crystals of the MAM L50A and K201A mutants, respectively. Molecular-replacement calculations suggest the presence of two molecules in the asymmetric unit for the MAM K201A mutant crystal, resulting in a V{sub M} of 5.0 Angstroms Da{sup -1} and a solvent content of 75%. Anmore » interpretable electron-density map for the MAM K201A mutant crystal was produced using the molecular-replacement method.« less

Authors:
; ; ;
Publication Date:
Research Org.:
Brookhaven National Laboratory (BNL) National Synchrotron Light Source
Sponsoring Org.:
Doe - Office Of Science
OSTI Identifier:
914016
Report Number(s):
BNL-78584-2007-JA
TRN: US200804%%430
DOE Contract Number:  
DE-AC02-98CH10886
Resource Type:
Journal Article
Resource Relation:
Journal Name: Acta Cryst. F; Journal Volume: 62
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; ALANINES; CHEMICAL ANALYSIS; CRYSTALLIZATION; DIFFRACTION; ELECTRON DENSITY; GENE MUTATIONS; MITOGENS; MUTAGENESIS; MUTANTS; MYCOPLASMA; RESOLUTION; SOLVENTS; SPACE GROUPS; national synchrotron light source

Citation Formats

Guo,Y., Li, Z., Van Vranken, S., and Li, H. A Single Point Mutation Changes in the Crystallization Behavior of Mycoplasma arthritidis-derived Mitogen. United States: N. p., 2006. Web. doi:10.1107/S1744309106003691.
Guo,Y., Li, Z., Van Vranken, S., & Li, H. A Single Point Mutation Changes in the Crystallization Behavior of Mycoplasma arthritidis-derived Mitogen. United States. doi:10.1107/S1744309106003691.
Guo,Y., Li, Z., Van Vranken, S., and Li, H. Sun . "A Single Point Mutation Changes in the Crystallization Behavior of Mycoplasma arthritidis-derived Mitogen". United States. doi:10.1107/S1744309106003691.
@article{osti_914016,
title = {A Single Point Mutation Changes in the Crystallization Behavior of Mycoplasma arthritidis-derived Mitogen},
author = {Guo,Y. and Li, Z. and Van Vranken, S. and Li, H.},
abstractNote = {Mycoplasma arthritidis-derived mitogen (MAM) functions as a conventional superantigen (SAg). Although recombinant MAM has been crystallized by the hanging-drop vapor-diffusion method, the crystals diffracted poorly to only 5.0 Angstroms resolution, with large unit-cell parameters a = 163.8, b = 93.0, c = 210.9 Angstroms, {beta} = 93.7 degrees in the monoclinic space group P2{sub 1}. Unit-cell content analysis revealed that as many as 24 molecules could be present in the asymmetric unit. Systematic alanine mutagenesis was applied in order to search for mutants that give crystals of better quality. Two mutants, L50A and K201A, were crystallized under the same conditions as wild-type MAM (MAM{sub wt}). Crystals of the L50A mutant are isomorphous with those of MAMwt, while a new crystal form was obtained for the K201 mutant, belonging to the cubic space group P4132 with unit-cell parameters a = b = c = 181.9 Angstroms. Diffraction data were collected to 3.6 and 2.8 Angstroms resolution from crystals of the MAM L50A and K201A mutants, respectively. Molecular-replacement calculations suggest the presence of two molecules in the asymmetric unit for the MAM K201A mutant crystal, resulting in a V{sub M} of 5.0 Angstroms Da{sup -1} and a solvent content of 75%. An interpretable electron-density map for the MAM K201A mutant crystal was produced using the molecular-replacement method.},
doi = {10.1107/S1744309106003691},
journal = {Acta Cryst. F},
number = ,
volume = 62,
place = {United States},
year = {Sun Jan 01 00:00:00 EST 2006},
month = {Sun Jan 01 00:00:00 EST 2006}
}