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Title: Crystallization and preliminary X-ray analysis of cytosolic α-mannosidase from Thermotoga maritima

Abstract

Cytosolic class II α-mannosidase from T. maritima (TM1851), a family 38 glycoside hydrolase, was crystallized. A diffraction data set was collected to 2.9 Å resolution. Class II α-mannosidase cleaves off α-1,2-, α-1,3- and α-1,6-mannose residues. In this paper, the crystallization and preliminary X-ray analysis of cytosolic class II α-mannosidase from Thermotoga maritima (TM1851), a family 38 glycoside hydrolase, is described. The crystal of recombinant TM1851 belongs to the C-centred monoclinic space group C2, with unit-cell parameters a = 244.7, b = 87.4, c = 166.6 Å, β = 124.7°. X-ray diffraction data were collected to a resolution of 2.9 Å.

Authors:
; ; ; ;  [1]
  1. Department of Biotechnology, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657 (Japan)
Publication Date:
OSTI Identifier:
22356269
Resource Type:
Journal Article
Resource Relation:
Journal Name: Acta Crystallographica. Section F; Journal Volume: 62; Journal Issue: Pt 2; Other Information: PMCID: PMC2150957; PMID: 16511275; PUBLISHER-ID: pu5120; OAI: oai:pubmedcentral.nih.gov:2150957; Copyright (c) International Union of Crystallography 2006; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United Kingdom
Language:
English
Subject:
75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY; CRYSTALLIZATION; CRYSTALS; RESOLUTION; SPACE GROUPS; X-RAY DIFFRACTION

Citation Formats

Nakajima, Masahiro, Fushinobu, Shinya, E-mail: asfushi@mail.ecc.u-tokyo.ac.jp, Imamura, Hiromi, Shoun, Hirofumi, and Wakagi, Takayoshi. Crystallization and preliminary X-ray analysis of cytosolic α-mannosidase from Thermotoga maritima. United Kingdom: N. p., 2006. Web. doi:10.1107/S1744309105042508.
Nakajima, Masahiro, Fushinobu, Shinya, E-mail: asfushi@mail.ecc.u-tokyo.ac.jp, Imamura, Hiromi, Shoun, Hirofumi, & Wakagi, Takayoshi. Crystallization and preliminary X-ray analysis of cytosolic α-mannosidase from Thermotoga maritima. United Kingdom. doi:10.1107/S1744309105042508.
Nakajima, Masahiro, Fushinobu, Shinya, E-mail: asfushi@mail.ecc.u-tokyo.ac.jp, Imamura, Hiromi, Shoun, Hirofumi, and Wakagi, Takayoshi. Wed . "Crystallization and preliminary X-ray analysis of cytosolic α-mannosidase from Thermotoga maritima". United Kingdom. doi:10.1107/S1744309105042508.
@article{osti_22356269,
title = {Crystallization and preliminary X-ray analysis of cytosolic α-mannosidase from Thermotoga maritima},
author = {Nakajima, Masahiro and Fushinobu, Shinya, E-mail: asfushi@mail.ecc.u-tokyo.ac.jp and Imamura, Hiromi and Shoun, Hirofumi and Wakagi, Takayoshi},
abstractNote = {Cytosolic class II α-mannosidase from T. maritima (TM1851), a family 38 glycoside hydrolase, was crystallized. A diffraction data set was collected to 2.9 Å resolution. Class II α-mannosidase cleaves off α-1,2-, α-1,3- and α-1,6-mannose residues. In this paper, the crystallization and preliminary X-ray analysis of cytosolic class II α-mannosidase from Thermotoga maritima (TM1851), a family 38 glycoside hydrolase, is described. The crystal of recombinant TM1851 belongs to the C-centred monoclinic space group C2, with unit-cell parameters a = 244.7, b = 87.4, c = 166.6 Å, β = 124.7°. X-ray diffraction data were collected to a resolution of 2.9 Å.},
doi = {10.1107/S1744309105042508},
journal = {Acta Crystallographica. Section F},
number = Pt 2,
volume = 62,
place = {United Kingdom},
year = {Wed Feb 01 00:00:00 EST 2006},
month = {Wed Feb 01 00:00:00 EST 2006}
}
  • Crystals of the peroxiredoxin domain of a larger natural hybrid protein from T. maritima were obtained which diffracted to 2.9 Å resolution on a synchrotron source. Thermotoga maritima contains a natural hybrid protein constituted of two moieties: a peroxiredoxin domain at the N-terminus and a nitroreductase domain at the C-terminus. The peroxiredoxin (Prx) domain has been overproduced and purified from Escherichia coli cells. The recombinant Prx domain, which is homologous to bacterial Prx BCP and plant Prx Q, folds properly into a stable protein that possesses biological activity. The recombinant protein was crystallized and synchrotron data were collected to 2.9more » Å resolution. The crystals belonged to the tetragonal space group I422, with unit-cell parameters a = b = 176.67, c = 141.20 Å.« less
  • T. maritima mannitol dehydrogenase has been crystallized in space group P2{sub 1}2{sub 1}2{sub 1} with a = 84.43, b = 120.61, c = 145.76 Å. The crystals diffracted to 3.3 Å resolution at the Canadian Light Source. Diffraction data have been collected from a crystal of Thermotoga maritima mannitol dehydrogenase at the Canadian Light Source. The crystal diffracted to 3.3 Å resolution and belongs to space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 83.43, b = 120.61, c = 145.76 Å. The structure is likely to be solved by molecular replacement.
  • T. maritima TrmFO was overexpressed, purified and crystallized. A diffraction data set was collected to a resolution of 2.6 Å. TrmFO, previously classified as GID, is a methyltransferase that catalyzes the formation of 5-methyluridine or ribothymidine (T) at position 54 in tRNA in some Gram-positive bacteria. To date, TrmFO is the only characterized tRNA methyltransferase that does not use S-adenosylmethionine as the methyl-group donor. Instead, the donor of the methyl group is N{sup 5},N{sup 10}-methylenetetrahydrofolate. The crystallization and preliminary X-ray crystallographic studies of TrmFO are reported here. The recombinant protein, cloned from Thermotoga maritima genomic DNA, was overproduced in Esherichiamore » coli and crystallized in 25%(v/v) PEG 4000, 100 mM NaCl and sodium citrate buffer pH 5.0 at 291 K using the hanging-drop vapor-diffusion method. The plate-shaped crystals diffracted to 2.6 Å and belong to the orthorhombic space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 79.94, b = 92.46, c = 127.20 Å.« less
  • The DNA repair enzyme Endonuclease IV from the thermophilic bacterium Thermotoga Maritima MSB8 (reference sequence: NC_000853) has been expressed in Escherichia coli and crystallized for X ray analysis. Thermotoga maritima Endonuclease IV is a 287 amino acid protein with 32% sequence identity to the Escherichia coli Endonuclease IV. The protein was purified to homogeneity and was crystallized using the sitting drop vapor diffusion method. The protein crystallized in the space group P61, with a composition of one biological molecule in the asymmetric unit corresponding to a Mathew s coefficient of 2.39 and a 47% solvent fraction. The unit cell parametersmore » for the crystals are a = 123.23 , b = 123.23 , c = 35.34 , = = 90 , = 120 . Microseeding and further optimization yielded crystals with an X ray diffraction limit of 2.4 . A single 70 data set was collected and processed resulting in an overall Rmerge and completeness of 9.5% and 99.3% respectively.« less
  • A thermostable esterase (EstA) from Thermotoga maritima was cloned and purified. Crystals of EstA and its selenomethionine derivative were grown and diffract to beyond 2.6 Å resolution at 100 K using synchrotron radiation. A predicted esterase (EstA) with an unusual new domain from the hyperthermophilic bacterium Thermotoga maritima has been cloned and overexpressed in Escherichia coli. The purified protein was crystallized by the hanging-drop vapour-diffusion technique in the presence of lithium sulfate and polyethylene glycol 8000. Selenomethionine-substituted EstA crystals were obtained under the same conditions and three different-wavelength data sets were collected to 2.6 Å resolution. The crystal belongs tomore » space group H32, with unit-cell parameters a = b = 130.2, c = 306.2 Å. There are two molecules in the asymmetric unit, with a V{sub M} of 2.9 Å{sup 3} Da{sup −1} and 58% solvent content.« less