A5020367412- Monoclonal and Polyclonal Antibodies Research
- Enzyme Structure and Function
- Glycosylation and Glycoproteins Research
- Protein Structure and Dynamics
- Protein purification and stability
- RNA and protein synthesis mechanisms
- Cell Adhesion Molecules Research
- Biochemical and Molecular Research
- T-cell and B-cell Immunology
- Enzyme Production and Characterization
- Viral Infectious Diseases and Gene Expression in Insects
- Peptidase Inhibition and Analysis
- Immunotherapy and Immune Responses
- Carbohydrate Chemistry and Synthesis
- Bacterial Genetics and Biotechnology
- Toxin Mechanisms and Immunotoxins
- Immune Response and Inflammation
- ATP Synthase and ATPases Research
- Synthesis and Characterization of Heterocyclic Compounds
- Biochemical and Structural Characterization
- Metabolism and Genetic Disorders
- Immune Cell Function and Interaction
- Hemoglobin structure and function
- Mass Spectrometry Techniques and Applications
- Polyamine Metabolism and Applications
Janssen (United States)
2013-2023
Janssen (Switzerland)
2015
Janssen (Belgium)
2012-2014
Biotechnology Institute
2001-2009
National Institute of Standards and Technology
2001-2005
Illinois Institute of Technology
2005
University of New Mexico
2005
Physical Sciences (United States)
2005
National Institutes of Health
1998-2004
University of York
1995-2004
MOLREP is an automated program for molecular replacement which utilizes effective new approaches in data processing and rotational translational searching. These include automatic choice of all parameters, scaling by Patterson origin peaks soft resolution cut-off. One the cornerstones original full-symmetry translation function combined with a packing function. Information from model already placed cell incorporated both functions. A number tests using experimental proved ability to find...
MOLREP is an automated program for molecular replacement that utilizes a number of original approaches to rotational and translational search data preparation. Since the first publication describing program, has acquired variety features include weighting X-ray models, multi-copy search, fitting model into electron density, structural superposition two models rigid-body refinement. The can run in fully automatic mode using optimized parameters calculated from input data.
The molecular-replacement method has been extended to a simultaneous search for multiple copies of the macromolecule in unit cell. central point this approach is construction multi-copy model from properly oriented monomers using special translation function. implemented program MOLREP and successfully tested experimental data.
D-amino acid oxidase is the prototype of FAD-dependent oxidases. It catalyses oxidation acids to corresponding alpha-ketoacids. The reducing equivalents are transferred molecular oxygen with production hydrogen peroxide. We have solved crystal structure complex benzoate, a competitive inhibitor substrate, by single isomorphous replacement and eightfold averaging. Each monomer formed two domains an overall topology similar that p-hydroxybenzoate hydroxylase. benzoate molecule lays parallel...
Protein aggregation is of great concern to pharmaceutical formulations and has been implicated in several diseases. We engineered an anti-IL-13 monoclonal antibody CNTO607 for improved solubility. Three structure-based engineering approaches were employed this study: (i) modifying the isoelectric point (pI), (ii) decreasing overall surface hydrophobicity (iii) re-introducing N-linked carbohydrate moiety within a complementarity-determining region (CDR) sequence. A mutant was identified with...
ABSTRACT To assess the state of art in antibody 3D modeling, 11 unpublished high‐resolution x‐ray Fab crystal structures from diverse species and covering a wide range antigen‐binding site conformations were used as benchmark to compare Fv models generated by seven structure prediction methodologies. The participants included: Accerlys Inc, Chemical Computer Group (CCG), Schrodinger, Jeff Gray's lab at John Hopkins University, Macromoltek, Astellas Pharma/Osaka University Prediction...
A blinded study to assess the state of art in three-dimensional structure modeling variable region (Fv) antibodies was conducted. Nine unpublished high-resolution x-ray Fab crystal structures covering a wide range antigen-binding site conformations were used as benchmark compare Fv models generated by four prediction methodologies. The methodologies included two homology strategies independently developed CCG (Chemical Computer Group) and Accerlys Inc, fully automated antibody servers: PIGS...
Microtubule-associated protein tau becomes abnormally phosphorylated in Alzheimer's disease and other tauopathies forms aggregates of paired helical filaments (PHF-tau). AT8 is a PHF-tau-specific monoclonal antibody that commonly used marker neuropathology because its recognition tau. Previous reports described the epitope to include pS202/pT205. Our studies support extend previous findings by also identifying pS208 as part binding epitope. We characterized phosphoepitope through both...
The use of consensus design to produce stable proteins has been applied numerous structures and classes proteins. Here, we describe the engineering novel FN3 domains from two different proteins, namely human fibronectin tenascin-C, as potential alternative scaffold biotherapeutics. resulting were found be robustly expressed in Escherichia coli, soluble highly stable, with melting temperatures 89 78°C, respectively. X-ray crystallography was used confirm that approach led a structure...
Some antibodies have a tendency to self-associate leading precipitation at relatively low concentrations. CNTO607, monoclonal antibody, precipitates irreversibly in phosphate-buffered saline concentrations above 13 mg/ml. Previous mutagenesis work based on the Fab crystal structure pinpointed three residue fragment heavy chain CDR-3, (99)FHW(100a), as an aggregation epitope that is anchored by two salt bridges. Biophysical characterization of variants reveals F99 and W100a, but not H100,...
To assess the state-of-the-art in antibody structure modeling, a blinded study was conducted. Eleven unpublished Fab crystal structures were used as benchmark to compare Fv models generated by seven prediction methodologies. In first round, each participant submitted three non-ranked complete for target. second CDR-H3 modeling performed context of correct environment provided with removed. this report we describe reference and present our assessment models. Some essential sources errors...
To support antibody therapeutic development, the crystal structures of a set 16 germline variants composed 4 different kappa light chains paired with heavy have been determined. All four antigen-binding fragments (Fabs) same complementarity-determining region (CDR) H3 that was reported in an earlier Fab structure. The structure analyses include comparisons overall structures, canonical CDRs and VH:VL packing interactions. CDR conformations for most part are tightly clustered, especially ones...
Single-chain fragment variable (scFv) domains play an important role in antibody-based therapeutic modalities, such as bispecifics, multispecifics and chimeric antigen receptor T cells or natural killer cells. However, scFv exhibit lower stability increased risk of aggregation due to transient dissociation ("breathing") inter-molecular reassociation the two (VL VH). We designed a novel strategy, referred stapling, that introduces disulfide bonds between linker minimize breathing. named...
The wealth of kinetic and structural information makes inorganic pyrophosphatases (PPases) a good model system to study the details enzymatic phosphoryl transfer. enzyme accelerates metal-complexed transfer 10 -fold: but how? Our structures yeast PPase product complex at 1.15 Å fluoride-inhibited 1.9 visualize active site in three different states: substrate-bound, immediate bound, relaxed bound. These span steps around chemical catalysis provide strong evidence that water molecule (O nu )...
Bacterial transport of many sugars, coupled to their phosphorylation, is carried out by the phosphoenolpyruvate (PEP):sugar phosphotransferase system and involves five phosphoryl group transfer reactions. Sugar translocation initiates with Mg(2+)-dependent phosphorylation enzyme I (EI) PEP. Crystals Escherichia coli EI were obtained mixing protein Mg(2+) PEP, followed oxalate, an inhibitor. The crystal structure reveals a dimeric where each subunit comprises three domains: domain that binds...
Glucosamine 6-phosphate synthase converts fructose-6P into glucosamine-6P or glucose-6P depending on the presence absence of glutamine. The isomerase activity is associated with a 40-kDa C-terminal domain, which has already been characterized crystallographically. Now three-dimensional structures complexes reaction product and transition state analog 2-amino-2-deoxyglucitol-6P have determined. Glucose-6P binds in cyclic form whereas an extended conformation. information ligand-protein...