Evidence map›Paper›PMID 25295172›Full record

ArticleIUCrJ2014

Expression, purification and crystallization of CTB-MPR, a candidate mucosal vaccine component against HIV-1.

Ho-Hsien Lee, Irene Cherni, HongQi Yu, Raimund Fromme, Jeffrey D Doran, Ingo Grotjohann, Michele Mittman, Shibom Basu, Arpan Deb, Katerina Dörner and 27 more

Open access · goldAbstract read
PubMed Full text at PMC Publisher
In one paragraph

Article in IUCrJ, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 5 papers.

0numbers the graph read from it
0cells of the map it votes in
5citing papers in PubMed
0.8field-weighted citation impact, top 31% of its field
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

5 citing papers in PubMed, 7 citations in OpenAlex.

  1. Article
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  4. Article
  5. Article
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

37 authors at 7 institutions in 2 countries.

Ho-Hsien LeeDepartment of Chemistry and Biochemistry, Arizona State University , PO Box 871604, Tempe, AZ 85287-1604, USA.
Irene CherniSchool of Life Sciences, Arizona State University , PO Box 874501, Tempe, AZ 85287-4501, USA ; Center for Infectious Diseases and Vaccinology, Biodesign Institute, Arizona State University , PO Box 874501, Tempe, AZ 85287-5401, USA.
HongQi YuDepartment of Chemistry and Biochemistry, Arizona State University , PO Box 871604, Tempe, AZ 85287-1604, USA.
Raimund FrommeDepartment of Chemistry and Biochemistry, Arizona State University , PO Box 871604, Tempe, AZ 85287-1604, USA.
Jeffrey D DoranSchool of Life Sciences, Arizona State University , PO Box 874501, Tempe, AZ 85287-4501, USA ; Center for Infectious Diseases and Vaccinology, Biodesign Institute, Arizona State University , PO Box 874501, Tempe, AZ 85287-5401, USA.
Ingo GrotjohannDepartment of Chemistry and Biochemistry, Arizona State University , PO Box 871604, Tempe, AZ 85287-1604, USA.
Michele MittmanSchool of Life Sciences, Arizona State University , PO Box 874501, Tempe, AZ 85287-4501, USA ; Center for Infectious Diseases and Vaccinology, Biodesign Institute, Arizona State University , PO Box 874501, Tempe, AZ 85287-5401, USA.
Shibom BasuDepartment of Chemistry and Biochemistry, Arizona State University , PO Box 871604, Tempe, AZ 85287-1604, USA.
Arpan DebSchool of Life Sciences, Arizona State University , PO Box 874501, Tempe, AZ 85287-4501, USA ; Center for Infectious Diseases and Vaccinology, Biodesign Institute, Arizona State University , PO Box 874501, Tempe, AZ 85287-5401, USA.
Katerina DörnerDepartment of Chemistry and Biochemistry, Arizona State University , PO Box 871604, Tempe, AZ 85287-1604, USA.
Andrew AquilaCenter for Free-Electron Laser Science, DESY , Notkestrasse 85, 22607 Hamburg, Germany.
Anton BartyCenter for Free-Electron Laser Science, DESY , Notkestrasse 85, 22607 Hamburg, Germany.
Sébastien BoutetLinac Coherent Light Source, SLAC National Accelerator Laboratory , 2575 Sand Hill Road, Menlo Park, CA 94025, USA.
Henry N ChapmanCenter for Free-Electron Laser Science, DESY , Notkestrasse 85, 22607 Hamburg, Germany ; University of Hamburg , Luruper Chaussee 149, 22761 Hamburg, Germany.
R Bruce DoakDepartment of Physics, Arizona State University , PO Box 871504, Tempe, AZ 85287-1504, USA.
Mark S HunterDepartment of Chemistry and Biochemistry, Arizona State University , PO Box 871604, Tempe, AZ 85287-1604, USA.
Daniel JamesDepartment of Physics, Arizona State University , PO Box 871504, Tempe, AZ 85287-1504, USA.
Richard A KirianCenter for Free-Electron Laser Science, DESY , Notkestrasse 85, 22607 Hamburg, Germany ; Department of Physics, Arizona State University , PO Box 871504, Tempe, AZ 85287-1504, USA.
Christopher KupitzDepartment of Chemistry and Biochemistry, Arizona State University , PO Box 871604, Tempe, AZ 85287-1604, USA.
Robert M LawrenceDepartment of Chemistry and Biochemistry, Arizona State University , PO Box 871604, Tempe, AZ 85287-1604, USA ; Center for Infectious Diseases and Vaccinology, Biodesign Institute, Arizona State University , PO Box 874501, Tempe, AZ 85287-5401, USA.
Haiguang LiuDepartment of Physics, Arizona State University , PO Box 871504, Tempe, AZ 85287-1504, USA.
Karol NassCenter for Free-Electron Laser Science, DESY , Notkestrasse 85, 22607 Hamburg, Germany ; University of Hamburg , Luruper Chaussee 149, 22761 Hamburg, Germany.
Ilme SchlichtingMax-Planck-Institut für medizinische Forschung , Jahnstrasse 29, 69120 Heidelberg, Germany.
Kevin E SchmidtDepartment of Physics, Arizona State University , PO Box 871504, Tempe, AZ 85287-1504, USA.
M Marvin SeibertLinac Coherent Light Source, SLAC National Accelerator Laboratory , 2575 Sand Hill Road, Menlo Park, CA 94025, USA.
Robert L ShoemanMax-Planck-Institut für medizinische Forschung , Jahnstrasse 29, 69120 Heidelberg, Germany.
John C H SpenceDepartment of Physics, Arizona State University , PO Box 871504, Tempe, AZ 85287-1504, USA.
Francesco StellatoCenter for Free-Electron Laser Science, DESY , Notkestrasse 85, 22607 Hamburg, Germany.
Uwe WeierstallDepartment of Physics, Arizona State University , PO Box 871504, Tempe, AZ 85287-1504, USA.
Garth J WilliamsLinac Coherent Light Source, SLAC National Accelerator Laboratory , 2575 Sand Hill Road, Menlo Park, CA 94025, USA.
Chunhong YoonCenter for Free-Electron Laser Science, DESY , Notkestrasse 85, 22607 Hamburg, Germany ; European XFEL GmbH , Albert-Einstein-Ring 19, 22761 Hamburg, Germany.
Dingjie WangDepartment of Physics, Arizona State University , PO Box 871504, Tempe, AZ 85287-1504, USA.
Nadia A ZatsepinDepartment of Physics, Arizona State University , PO Box 871504, Tempe, AZ 85287-1504, USA.
Brenda G HogueSchool of Life Sciences, Arizona State University , PO Box 874501, Tempe, AZ 85287-4501, USA ; Center for Infectious Diseases and Vaccinology, Biodesign Institute, Arizona State University , PO Box 874501, Tempe, AZ 85287-5401, USA.
Nobuyuki MatobaSchool of Life Sciences, Arizona State University , PO Box 874501, Tempe, AZ 85287-4501, USA ; Center for Infectious Diseases and Vaccinology, Biodesign Institute, Arizona State University , PO Box 874501, Tempe, AZ 85287-5401, USA.
Petra FrommeDepartment of Chemistry and Biochemistry, Arizona State University , PO Box 871604, Tempe, AZ 85287-1604, USA.
Tsafrir S MorSchool of Life Sciences, Arizona State University , PO Box 874501, Tempe, AZ 85287-4501, USA ; Center for Infectious Diseases and Vaccinology, Biodesign Institute, Arizona State University , PO Box 874501, Tempe, AZ 85287-5401, USA.
Arizona State University · USUniversität Hamburg · DEMax Planck Institute for Medical Research · DESLAC National Accelerator Laboratory · USCenter for Free-Electron Laser Science · DEEuropean X-Ray Free-Electron Laser · DELinac Coherent Light Source · US

Funding

Production CoreU19AI062150 · NIAID · ARIZONA STATE UNIVERSITY-TEMPE CAMPUS · PI MASON, HUGH · 2004 to 2008
$7.2M
Center for the Rational Design of Membrane Protein CrystallographyU54GM094599 · NIGMS · ARIZONA STATE UNIVERSITY-TEMPE CAMPUS · PI FROMME, PETRA · 2010 to 2014
$6.8M
Femtosecond nano-crystallography of membrane proteinsR01GM095583 · NIGMS · ARIZONA STATE UNIVERSITY-TEMPE CAMPUS · PI FROMME, PETRA · 2010 to 2023
$4.3M
Expression of Decontructed HIV-1 Virus-Like Particles in Bioengineered PlantsR03AI073157 · NIAID · UNIVERSITY OF LOUISVILLE · PI MATOBA, NOBUYUKI · 2007 to 2008
$148k
NIAID NIH HHS R03 AI073157NIAID NIH HHS U19 AI062150NIGMS NIH HHS R01 GM095583NIGMS NIH HHS U54 GM094599
6 · The paper itself

Abstract

CTB-MPR is a fusion protein between the B subunit of cholera toxin (CTB) and the membrane-proximal region of gp41 (MPR), the transmembrane envelope protein of Human immunodeficiency virus 1 (HIV-1), and has previously been shown to induce the production of anti-HIV-1 antibodies with antiviral functions. To further improve the design of this candidate vaccine, X-ray crystallography experiments were performed to obtain structural information about this fusion protein. Several variants of CTB-MPR were designed, constructed and recombinantly expressed in Escherichia coli. The first variant contained a flexible GPGP linker between CTB and MPR, and yielded crystals that diffracted to a resolution of 2.3 Å, but only the CTB region was detected in the electron-density map. A second variant, in which the CTB was directly attached to MPR, was shown to destabilize pentamer formation. A third construct containing a polyalanine linker between CTB and MPR proved to stabilize the pentameric form of the protein during purification. The purification procedure was shown to produce a homogeneously pure and monodisperse sample for crystallization. Initial crystallization experiments led to pseudo-crystals which were ordered in only two dimensions and were disordered in the third dimension. Nanocrystals obtained using the same precipitant showed promising X-ray diffraction to 5 Å resolution in femtosecond nanocrystallography experiments at the Linac Coherent Light Source at the SLAC National Accelerator Laboratory. The results demonstrate the utility of femtosecond X-ray crystallography to enable structural analysis based on nano/microcrystals of a protein for which no macroscopic crystals ordered in three dimensions have been observed before.

Indexed as

cholera toxin B subunitcrystallizationfemtosecond nanocrystallographyfree-electron lasersgp41HIV-1membrane-proximal regionX-ray crystallography

Identifiers

PMID25295172
PMCPMC4174873
OpenAlexW2046380801

What OpenQuestion holds

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Registered trials

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.