ArticleProtein science : a publication of the Protein Society2003
Three-dimensional crystallization of the Escherichia coli glycerol-3-phosphate transporter: a member of the major facilitator superfamily.
Article in Protein science : a publication of the Protein Society, 2003. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 39 papers.
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39 citing papers in PubMed, 96 citations in OpenAlex.
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- Reconciling contradictory findings: Glucose transporter 1 (GLUT1) functions as an oligomer of allosteric, alternating access transporters.The Journal of biological chemistry · 2017Article
- Mycobacterium tuberculosis Major Facilitator Superfamily Transporters.The Journal of membrane biology · 2017Review
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- Quantification of Detergents Complexed with Membrane Proteins.Scientific reports · 2017Article
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- Structural Insights into the Transport Mechanism of the Human Sodium-dependent Lysophosphatidylcholine Transporter MFSD2A.The Journal of biological chemistry · 2016Article
- Glucose Transporters in Cardiac Metabolism and Hypertrophy.Comprehensive Physiology · 2015Review
- Deletion of the 2-acyl-glycerophosphoethanolamine cycle improve glucose metabolism in Escherichia coli strains employed for overproduction of aromatic compounds.Microbial cell factories · 2015Article
- Isolation and characterisation of transport-defective substrate-binding mutants of the tetracycline antiporter TetA(B).Biochimica et biophysica acta · 2015Article
- Atomic-level characterization of transport cycle thermodynamics in the glycerol-3-phosphate:phosphate antiporter.Nature communications · 2015Article
- Multidrug efflux pumps from Enterobacteriaceae, Vibrio cholerae and Staphylococcus aureus bacterial food pathogens.International journal of environmental research and public health · 2015Review
- Production of functional human vitamin A transporter/RBP receptor (STRA6) for structure determination.PloS one · 2015Article
- Membrane proteins, detergents and crystals: what is the state of the art?Acta crystallographica. Section F, Structural biology communications · 2014Review
- Variation of the detergent-binding capacity and phospholipid content of membrane proteins when purified in different detergents.Biophysical journal · 2014Article
- A decade of crystallization drops: crystallization of the cbb3 cytochrome c oxidase from Pseudomonas stutzeri.Protein science : a publication of the Protein Society · 2014Article
- Opening the black box: revealing the molecular basis of thyroid hormone transport.Endocrinology · 2013Article
- Rapid expression screening of eukaryotic membrane proteins in Pichia pastoris.Protein science : a publication of the Protein Society · 2013Article
- A fluorescence-detection size-exclusion chromatography-based thermostability assay for membrane protein precrystallization screening.Structure (London, England : 1993) · 2012Article
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8 authors at 1 institution in 1 country.
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Abstract
Here we report the successful three-dimensional crystallization of GlpT, the glycerol-3-phosphate transporter from Escherichia coli inner membrane. GlpT possesses 12 transmembrane alpha-helices and is a member of the major facilitator superfamily. It mediates the exchange of glycerol-3-phosphate for inorganic phosphate across the membrane. Approximately 20 phospholipid molecules per protein, identified as negatively charged phosphatidylethanolamine, phosphatidylglycerol, and cardiolipin, were required for the monodispersity of purified GlpT. Analytical size-exclusion chromatography proved to be efficient in identifying detergents for GlpT monodispersity. Nine such detergents were later used for GlpT crystallization. Screening for crystal nucleation was carried out with a variety of polyethylene glycols as the precipitant over a wide pH range. Subsequent identification of a rigid protein core by limited proteolysis and mass spectroscopy resulted in better-ordered crystals. These crystals exhibited order to 3.7 A resolution in two dimensions. However, the stacking in the third dimension was partially disordered. This stacking problem was overcome by using a detergent mixture and manipulating the ionic interactions in the crystallization solution. The resulting GlpT crystals diffracted isotropically to 3.3 A resolution and were suitable for structure determination by X-ray crystallography.
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