Proteolysis of the type III glutamine synthetase from Bacteroides fragilis causes expedient crystal-packing rearrangements. 2011

Jason van Rooyen, and Hassan Belrhali, and Valarie Abratt, and B Trevor Sewell
Electron Microscope Unit, Department of Molecular and Cell Biology, University of Cape Town, South Africa. jason@science.uct.ac.za

This work details the intentional modifications that led to the first structure of a type III glutamine synthetase enzyme (GSIII). This approach followed the serendipitous discovery of digestion caused by an extracellular protease from a contaminating bacterium, Pseudomonas fluorescens. The protease only cleaves the GSIII protein at a single site, leaving the oligomer intact but allowing the protein to crystallize in a different space group. This transition from space group P1 to space group C222(1) is accompanied by improved growth characteristics, more reproducible diffraction and enhanced mechanical stability. The crystallographic analyses presented here provide the structural basis of the altered molecular packing in the full-length and digested crystal forms and suggest modifications for future structural studies.

UI MeSH Term Description Entries
D008958 Models, Molecular Models used experimentally or theoretically to study molecular shape, electronic properties, or interactions; includes analogous molecules, computer-generated graphics, and mechanical structures. Molecular Models,Model, Molecular,Molecular Model
D008969 Molecular Sequence Data Descriptions of specific amino acid, carbohydrate, or nucleotide sequences which have appeared in the published literature and/or are deposited in and maintained by databanks such as GENBANK, European Molecular Biology Laboratory (EMBL), National Biomedical Research Foundation (NBRF), or other sequence repositories. Sequence Data, Molecular,Molecular Sequencing Data,Data, Molecular Sequence,Data, Molecular Sequencing,Sequencing Data, Molecular
D011487 Protein Conformation The characteristic 3-dimensional shape of a protein, including the secondary, supersecondary (motifs), tertiary (domains) and quaternary structure of the peptide chain. PROTEIN STRUCTURE, QUATERNARY describes the conformation assumed by multimeric proteins (aggregates of more than one polypeptide chain). Conformation, Protein,Conformations, Protein,Protein Conformations
D005974 Glutamate-Ammonia Ligase An enzyme that catalyzes the conversion of ATP, L-glutamate, and NH3 to ADP, orthophosphate, and L-glutamine. It also acts more slowly on 4-methylene-L-glutamate. (From Enzyme Nomenclature, 1992) EC 6.3.1.2. Glutamine Synthetase,Glutamate Ammonia Ligase (ADP),Glutamate Ammonia Ligase,Ligase, Glutamate-Ammonia,Synthetase, Glutamine
D001441 Bacteroides fragilis Gram-negative bacteria occurring in the lower intestinal tracts of man and other animals. It is the most common species of anaerobic bacteria isolated from human soft tissue infections.
D018360 Crystallography, X-Ray The study of crystal structure using X-RAY DIFFRACTION techniques. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed) X-Ray Crystallography,Crystallography, X Ray,Crystallography, Xray,X Ray Crystallography,Xray Crystallography,Crystallographies, X Ray,X Ray Crystallographies
D019032 Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization A mass spectrometric technique that is used for the analysis of large biomolecules. Analyte molecules are embedded in an excess matrix of small organic molecules that show a high resonant absorption at the laser wavelength used. The matrix absorbs the laser energy, thus inducing a soft disintegration of the sample-matrix mixture into free (gas phase) matrix and analyte molecules and molecular ions. In general, only molecular ions of the analyte molecules are produced, and almost no fragmentation occurs. This makes the method well suited for molecular weight determinations and mixture analysis. Laser Desorption-Ionization Mass Spectrometry, Matrix-Assisted,MALD-MS,MALDI,Mass Spectrometry, Matrix-Assisted Laser Desorption-Ionization,Mass Spectroscopy, Matrix-Assisted Laser Desorption-Ionization,Matrix-Assisted Laser Desorption-Ionization Mass Spectrometry,Spectroscopy, Mass, Matrix-Assisted Laser Desorption-Ionization,MALDI-MS,MS-MALD,SELDI-TOF-MS,Surface Enhanced Laser Desorption Ionization Mass Spectrometry,Laser Desorption Ionization Mass Spectrometry, Matrix Assisted,MALDI MS,Mass Spectrometry, Matrix Assisted Laser Desorption Ionization,Mass Spectroscopy, Matrix Assisted Laser Desorption Ionization,Matrix Assisted Laser Desorption Ionization Mass Spectrometry

Related Publications

Jason van Rooyen, and Hassan Belrhali, and Valarie Abratt, and B Trevor Sewell
October 1986, Journal of general microbiology,
Jason van Rooyen, and Hassan Belrhali, and Valarie Abratt, and B Trevor Sewell
December 1989, Journal of general microbiology,
Jason van Rooyen, and Hassan Belrhali, and Valarie Abratt, and B Trevor Sewell
May 1998, Molecular & general genetics : MGG,
Jason van Rooyen, and Hassan Belrhali, and Valarie Abratt, and B Trevor Sewell
January 1972, Cold Spring Harbor symposia on quantitative biology,
Jason van Rooyen, and Hassan Belrhali, and Valarie Abratt, and B Trevor Sewell
January 1993, Journal of general microbiology,
Jason van Rooyen, and Hassan Belrhali, and Valarie Abratt, and B Trevor Sewell
July 1996, Structure (London, England : 1993),
Jason van Rooyen, and Hassan Belrhali, and Valarie Abratt, and B Trevor Sewell
October 2003, The Journal of biological chemistry,
Jason van Rooyen, and Hassan Belrhali, and Valarie Abratt, and B Trevor Sewell
January 1976, Journal of molecular biology,
Jason van Rooyen, and Hassan Belrhali, and Valarie Abratt, and B Trevor Sewell
April 1995, Archives of biochemistry and biophysics,
Jason van Rooyen, and Hassan Belrhali, and Valarie Abratt, and B Trevor Sewell
April 1982, European journal of biochemistry,
Copied contents to your clipboard!