PEPTIDE-CELL INTERACTIONS IN SACCHAROMYCES CEREVISIAE
R Fred
University Of Tennessee Knoxvillecity: Knoxville country: United States (us)
Grant 5R01GM022087-34 from National Institute Of General Medical Sciences
Abstract: G protein-coupled receptors (GPCRs) comprise the largest family of eukaryotic membrane proteins and are among the most widely-studied biological molecules due to their role as targets for many drugs used in human medicine and their involvement in a multitude of human pathologies. An understanding of GPCR activation, structure, and regulation will facilitate the design and synthesis of drugs to treat many diseases. Despite intense and sustained efforts by many scientists over the last twenty-five years, there remain large gaps in our knowledge of the structure and function of GPCRs. The goals of this proposal are to use the S. cerevisiae pheromone alpha-factor and its GPCR receptor Ste2p to develop approaches that can be used to learn about how activated receptors transmit a signal to intracellular G proteins, to acquire information on receptor structure in the resting and ligand-activated state, and to learn how GPCR signaling is regulated. Specifically, we will conduct experiments to (i) reveal the conformational changes in extracellular and intracellular domains of Ste2p that are triggered by agonist binding or constitutive mutation and define interactions between receptor and G protein, (ii) elucidate the three-dimensional structure of large fragments of GPCRs alone and as part of a reconstituted receptor, and (iii) study the regulation of signal transduction by identifying proteins that interact with the carboxyl-terminus of Ste2p and examine the conformational changes in the C-terminus induced by these interactions. To accomplish these goals we will use molecular biological, biochemical, and biophysical approaches including substituted cysteine accessibility mutagenesis, unnatural amino acid replacement, native chemical ligation, bioluminescent resonance energy transfer, and multidimensional nuclear magnetic resonance spectroscopy. Success in these studies should provide an understanding of GPCR structure and molecular mechanisms of activation, as well as new experimental tools that can be used by investigators studying this important group of human receptors. PUBLIC HEALTH RELEVANCE Peptide hormones control many essential biological processes in all organisms and tissues. Mutations in genes encoding hormone receptors lead to many pathological conditions in humans, and G protein-coupled receptors (GPCRs) are the target of a majority of pharmacological agents used in medicine today. An understanding of mechanism of action of GPCRs, the structure of these integral membrane proteins, and how signal transduction is regulated will fill fundamental gaps in our knowledge to facilitate the design and synthesis of drugs to treat many diseases
Keywords: Agonist; Amino Acids; base; Binding (Molecular Function); Binding Proteins; Binding Sites; Biochemical; Biological; Biological Process; Cell Communication; Chemicals; crosslink; Cysteine; design; Disease; drug synthesis; Drug usage; Energy Transfer; extracellular; Family; G Protein-Coupled Receptor Signaling; G-Protein-Coupled Receptors; G-substrate; Genes; Goals; GTP-Binding Proteins; Health; Heteronuclear NMR; Hormone Receptor; Human; Human Pathology; Integral Membrane Protein; Knowledge; Laboratories; Lead; Learning; Ligand Binding; Ligands; Ligation; Medicine; Membrane Proteins; Methods; Molecular; Molecular Biology; Molecular Structure; Mutagenesis; Mutation; NMR Spectroscopy; Organism; peptide hormone; Peptides; Phase; Pheromone; pheromone alpha-factor; photoactivation; protein activation; protein protein interaction; Proteins; Published Comment; receptor; Receptor Activation; receptor binding; reconstitution; Regulation; Research; Research Personnel; research study; Resolution; response; Rest; Role; Saccharomyces cerevisiae; Scientist; Signal Transduction; Solutions; Structure; Study Section; success; three dimensional structure; Tissues; tool; Transmembrane Domain
Project start date: 1978-05-01
Project end date: 2012-07-31
Budget start date: 1-AUG-2011
Budget end date: 31-JUL-2012
5R01GM022087-34 (2011): $660495
Sponsored Links Excellgen http://Excellgen.com
Grants awarded to R Fred
COMPETENCIES TRAINING &ASSESSMENT IN LABORATORYMEDICINE USINGPATIENTSIMULATIONS
R Fred
University Of Iowacity: Iowa City country: United States (us)
Grant 5R01LM009121-04 from National Library Of Medicine
Abstract: This proposal is designed to develop and evaluate a novel web-based model for computer assisted patient simulation that can be used for performance-based competency assessment and education. This new model will emulate patient care by the un-prompted, sequential decision making process of ordering, advance to the next encounter, and receiving and evaluating results; proceeding through several encounters, until the trainee decides to make a definitive diagnostic decision. As we develop this model, we will also evaluate whether a novel combined expert response/expert consensus scoring ajgorithm based on expert responses, augmented with evidence-based expert consensus building, conducted via web communication, provides better reliability and validity than expert response generated keys without the consensus component, or by global ratings. Reliability and validity of the simulation model and scoring key for competency assessment will be tested in the domain of clinical laboratory medicine. The simulation model and scoring key will be developed from a web-based simulation currently used in the pathology course at the University of Iowa (PathCAPS), which will be significantly upgraded in a Perl CGI scripted MySQL database. This newly developed simulation will assume the acronym LabCAPS. The evaluation research will be carried out in medical school courses at the University of Iowa an in laboratory medicine (clinical pathology) residency training programs at the University of Iowa Hospitals and Clinics and elsewhere. The educational goal of the simulation will be to foster evidence-based use of laboratory resources, and a reduction in unnecessary expenses. A long-term goal will be to develop a comprehensive web enabled dataset of LabCAPS simulations that can be used to reliably measure patient care competencies of medical trainees throughout the United States. With a successful outcome, the technology and scoring techniques developed by this grant can significantly improve the validity of testing in a number of areas in medical education. Because of validity concern with multiple-choice items, there has been widespread interest in using performance assessment methods. But performance assessments, as currently conceptualized, are extremely expensive to develop and score. This grant investigates methodologies designed to significantly reduce the cost of both development and scoring
Keywords: acronyms; Algorithms; Area; base; Characteristics; Clinical; clinical data repository; clinical data warehouse; Clinical Pathology; Clinics and Hospitals; Clinics or Hospitals; college; Competence; computer aided; Computer Assisted; computer program/software; Computer Programs; Computer software; Consensus; cost; cost effectiveness; Data; Data Banks; Data Bases; data repository; Data Set; Databank, Electronic; Databanks; Database, Electronic; Databases; Dataset; Decision Making; design; designing; Development; Diagnosis; Diagnostic; Education; Educational aspects; Evaluation; Evaluation Research; evidence base; Expenditure; Fostering; Goals; Grant; improved; Individual; interest; Internet; Investigators; Iowa; Laboratories; Measures; Medical; Medical Education; medical schools; Medical Students; Medicine; meetings; Method LOINC Axis 6; Methodology; Methods; Methods and Techniques; Methods, Other; Modeling; models and simulation; novel; On-Line Systems; online computer; Online Systems; Outcome; Pathology; Patient Care; Patient Care Delivery; Patient Simulation; Patients; Performance; Personal Satisfaction; Process; programs; Programs (PT); Programs [Publication Type]; relational database; Research; Research Personnel; Research Resources; Researchers; Residencies; Resources; response; Science of Medicine; simulation; Software; Techniques; Technology; Testing; Therapeutic; Training; Training Programs; United States; Universities; Validity and Reliability; web; web based; Web communication; web interface; web-enabled; well-being; world wide web; WWW
Project start date: 2007-03-01
Project end date: 2011-08-28
Budget start date: 1-MAR-2010
Budget end date: 28-AUG-2011
PFA/PA: PA-06-094
5R01LM009121-04 (2010): $257851
PEPTIDE-CELL INTERACTIONS IN SACCHAROMYCES CEREVISIAE
R Fred
University Of Tennessee Knoxvillecity: Knoxville country: United States (us)
Grant 3R01GM022087-33S1 from National Institute Of General Medical Sciences
Abstract: G protein-coupled receptors (GPCRs) comprise the largest family of eukaryotic membrane proteins and are among the most widely-studied biological molecules due to their role as targets for many drugs used in human medicine and their involvement in a multitude of human pathologies. An understanding of GPCR activation, structure, and regulation will facilitate the design and synthesis of drugs to treat many diseases. Despite intense and sustained efforts by many scientists over the last twenty-five years, there remain large gaps in our knowledge of the structure and function of GPCRs. The goals of this proposal are to use the S. cerevisiae pheromone alpha-factor and its GPCR receptor Ste2p to develop approaches that can be used to learn about how activated receptors transmit a signal to intracellular G proteins, to acquire information on receptor structure in the resting and ligand-activated state, and to learn how GPCR signaling is regulated. Specifically, we will conduct experiments to (i) reveal the conformational changes in extracellular and intracellular domains of Ste2p that are triggered by agonist binding or constitutive mutation and define interactions between receptor and G protein, (ii) elucidate the three-dimensional structure of large fragments of GPCRs alone and as part of a reconstituted receptor, and (iii) study the regulation of signal transduction by identifying proteins that interact with the carboxyl-terminus of Ste2p and examine the conformational changes in the C-terminus induced by these interactions. To accomplish these goals we will use molecular biological, biochemical, and biophysical approaches including substituted cysteine accessibility mutagenesis, unnatural amino acid replacement, native chemical ligation, bioluminescent resonance energy transfer, and multidimensional nuclear magnetic resonance spectroscopy. Success in these studies should provide an understanding of GPCR structure and molecular mechanisms of activation, as well as new experimental tools that can be used by investigators studying this important group of human receptors. PUBLIC HEALTH RELEVANCE Peptide hormones control many essential biological processes in all organisms and tissues. Mutations in genes encoding hormone receptors lead to many pathological conditions in humans, and G protein-coupled receptors (GPCRs) are the target of a majority of pharmacological agents used in medicine today. An understanding of mechanism of action of GPCRs, the structure of these integral membrane proteins, and how signal transduction is regulated will fill fundamental gaps in our knowledge to facilitate the design and synthesis of drugs to treat many diseases
Keywords: Agonist; Amino Acids; base; Binding (Molecular Function); Binding Proteins; Binding Sites; Biochemical; Biological; Biological Process; Cell Communication; Chemicals; crosslink; Cysteine; design; Disease; drug synthesis; Drug usage; Energy Transfer; extracellular; Family; G Protein-Coupled Receptor Signaling; G-Protein-Coupled Receptors; Genes; Goals; GTP-Binding Proteins; Heteronuclear NMR; Hormone Receptor; Human; Human Pathology; Integral Membrane Protein; Knowledge; Laboratories; Lead; Learning; Ligand Binding; Ligands; Ligation; Medicine; Membrane Proteins; Methods; Molecular; Molecular Biology; Molecular Structure; Mutagenesis; Mutation; NMR Spectroscopy; Organism; peptide hormone; Peptides; Phase; Pheromone; pheromone alpha-factor; photoactivation; protein activation; protein protein interaction; Proteins; public health relevance; Published Comment; receptor; Receptor Activation; receptor binding; reconstitution; Regulation; Research; Research Personnel; research study; Resolution; response; Rest; Role; Saccharomyces cerevisiae; Scientist; Signal Transduction; Solutions; Structure; Study Section; success; three dimensional structure; Tissues; tool; Transmembrane Domain
Project start date: 2010-09-01
Project end date: 2012-08-31
Budget start date: 1-SEP-2010
Budget end date: 31-AUG-2012
3R01GM022087-33S1 (2010): $284180