1985 — 1994 |
Marchase, Richard Banfield |
R01Activity Code Description: To support a discrete, specified, circumscribed project to be performed by the named investigator(s) in an area representing his or her specific interest and competencies. |
Glycoproteins Containing Phosphoglucose in Neural Retina @ University of Alabama At Birmingham
We have recently described a cytoplasmic phosphoglycoprotein of 62 kDa (pgp62) that appears to exist in two states, one containing solely an O- lined Man disaccharide and the other containing, in addition, phosphodiester-linked Glc. We have also described the enzymes responsible for the removal and addition of the glc-1-P. Our data support the hypothesis that pgp62 is intimately involved in the mechanism leading to synaptic vesicle release that the removal and addition of the Glc-1-P is an integral part of the exocytic process. We intend to critically test this hypothesis and to determine if the finding can be exploited to provide a high-resolution anatomical marker for active pre-synaptic terminals of neurons in the retina and lateral geniculate nucleus. The specific aims of this proposal are: 1.To continue the characterization of pgp62 and the enzymes involved in its regulation. This includes preparing antibodies specific for mammalian pgp62, determining its subcellular topography,k and cloning and sequencing it. In addition, the two enzymes that appear to be involved in its regulation, glc-1-P phosphodiesterase and glc phosphotransferase, will continue to be studied. 2.To test the hypothesis that synaptic vesicle release is accompanied by turnover of glc-1-P on pgp62 and to determine what the physiological significance of this modification is. This includes continuing biochemical characterization of glucose and phosphate metabolism in PC-12 cells and in synaptosomes and the development of an assay with permeabilized cells or synaptosomes that will allow us to assess the roles of isolated proteins, antibodies, and inhibitory factors n the secretory mechanism. 3.To determine in intact neurons the effects of stimulation on incorporation into macromolecules from labeled glc or 2-deoxyglucose and to determine if such differences can be exploited to develop a high-resolution marker for active pre-synaptic terminals in the retina and lateral geniculate nucleus.
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1 |
1985 |
Marchase, Richard Banfield |
R01Activity Code Description: To support a discrete, specified, circumscribed project to be performed by the named investigator(s) in an area representing his or her specific interest and competencies. |
Phosphoglucosyltranferase and Subcellular Localization
A requisite for the organizational complexity of eucaryotic cells is the efficient delivery of polypeptides from their sites of synthesis to the eventual sites of their function. The research this proposal addresses is a heretofore unexamined component of this problem, the mechanism by which a subclass of peripheral cell-surface glycoproteins are localized to the external surface of a cells' plasma membrane. We have recently completed two studies, the first of which determined that in embryonic chick neural retina glycoproteins that were recognized by the cell-surface protein ligatin contained high mannose-type oligosaccharides terminating in phosphodiester-linked glucose residues. The second described the presence of a phosphoglucosyltransferase responsible for the addition of those terminal residues. The hypothesis we propose to investigate suggests that this phosphoglucosyltransferase plays a salient role in the coding of its acceptor proteins for their subsequent transport to the cell surface. The proteins then remain at the cell surface through association with ligatin. We propose to address the following specific aims: 1) to identify the endogenous acceptor proteins of the phosphoglucosyltransferase in embryonic chick neural retina. We will compare polyacrylamide gels of the endogenous acceptor proteins with gels obtained from metabolically 32p-labelled cell-surface proteins associated with ligatin. We will also determine the ability of endogenous acceptor proteins to bind to ligatin affinity columns; 2) To purify and characterize the phosphoglucosyltransferase. In particular we plan to determine if glycoproteins destined for the cell surface are preferentially recognized by the transferase, and if so, to determine if intact tertiary structure of the acceptors is necessary of optimal activity; 3) To examine intracellular membrane fractions for an intracellular receptor for phosphoglucosyl-terminating oligosaccharides. Such a receptor could be necessary for the transport of the acceptor proteins to the cell surface; 4) To localize using both subcellular fraction techniques and electron microscope autoradiography the intracellular localization of the phosphoglucosyltransferase and to determine the route its acceptor proteins take as they are transported to the cell surface.
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0.97 |
1993 — 1999 |
Marchase, Richard Friedlander, Michael (co-PI) [⬀] |
N/AActivity Code Description: No activity code was retrieved: click on the grant title for more information |
Cellular and Molecular Neurobiology Graduate Training Program @ University of Alabama At Birmingham
20 faculty from 3 Departments (Cell Biology, Physiology and Biophysics, Psychology) are collaborating to provide a training program in cellular and molecular neuroscience. Over its course, this award will provide support for about 12 students. A major innovation in the training program is an intensive, 12 week summer laboratory course for students at the end of their first year of graduate school. The course consists of six 2-week modules in electrophysiology, developmental neurobiology, microscopy and imaging, second messengers and neurotransmitters, protein biochemistry, and molecular biology.
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0.915 |
2000 — 2003 |
Marchase, Richard Banfield |
R01Activity Code Description: To support a discrete, specified, circumscribed project to be performed by the named investigator(s) in an area representing his or her specific interest and competencies. |
Ca Influx Factor-- Structure/Function &Role in Diabetes @ University of Alabama At Birmingham
Numerous signal transduction pathways are dependent upon agonist-induced increases in cytoplasmic free Ca2+. The initial phase of this response is often due to the generation of inositol 1,4,5-trisphosphate and a subsequent release of Ca2+ from the endoplasmic reticulum (ER). The depletion of ER Ca2+ stores results in an influx of extracellular Ca2+ into the cytoplasm, a process termed capacitative or store-operated Ca2+ entry (SOCE). SOCE is implicated in a remarkable array of biological processes, including T cell activation, the secretion of insulin, vasoconstriction, and heart development. One hypothesis that has emerged to explain the transmission of information regarding ER Ca2+ depletion to the plasma membrane proposes a critical role for a novel diffusible messenger molecule, Ca2+ influx factor (ClF). In the last year data that have been gathered by this group of investigators provide strong support for the conclusion that CIF exists, that it is synthesized upon ER Ca2+ store depletion, and that it is responsible for activation of two distinct store-operated plasma membrane Ca2+ channels. Imaging and patch-clamp techniques for CIF's function have provided unique and independently verified data on mammalian CIF. In addition, this group has determined that Saccharomyces cerevisiae that are genetically deficient in an organellar Ca2+ ATPase also make a CIF that to date is indistinguishable from its mammalian counterpart. This inexpensive and plentiful source of material, the purification schemes for CIF that are well underway, and the assays that have been developed place this group in a unique position to purify and structurally characterize this novel signaling molecule. Experiments also will be performed to define the biosynthetic pathway for generating CIF upon ER Ca2+ store depletion, and to define the mechanism by which CIF activates the two plasma membrane Ca2+ + channels that contribute to SOCE. The information that we gain will be applied to a major health problem, diabetes, which mischaracterized by an underlying impairment of SOCE. The hyperglycemia-induced activation of the hexosamine biosynthetic pathway and/or protein kinase C will be investigated as possible regulators of SOCE. The described approach will involve a cross-disciplinary, multi-investigator initiative that applies biologic, biochemical, and structural procedures to this problem. Successful completion of these studies will provide pivotal information on SOCE and may define novel therapeutic targets for diabetes.
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1 |
2003 — 2006 |
Marchase, Richard Banfield |
UC6Activity Code Description: To provide financial support, substantial programmatic involvement and technical assistance for designing, constructing and commissioning of laboratories to be used as extramural facilities for bio-defense and emerging infectious diseases research. |
Construction/Seblab/Regional Biocontainment Laboratory @ University of Alabama At Birmingham
DESCRIPTION (provided by applicant): The University of Alabama at Birmingham (UAB) proposes to construct on its campus the Southeast Biocontainment Laboratory Alabama Birmingham (SEBLAB). This 41,060 gsf Regional Biocontainment Laboratory (RBL) will require $15.9 million of NIAID funding to be matched with $5.3 million from UAB. UAB is one of six institutions leading the Southeastern Regional Center for Excellence in Emerging Infections and Biodefense, the focus of biodefense efforts for the eight-state Region IV. UAB is an outstanding choice for an RBL for several reasons. These include the experience and complementary biodefense capabilities of UAB's affiliate, Southern Research Institute, the national leadership role that the UAB Center for Disaster Preparedness has taken in biodefense planning and training, and exceptional core facilities. Most importantly, UAB investigators have distinguished records in virology, bacterial pathogenesis, immunology, and vaccine development. Many of these investigators, as well as new recruits, are now bringing their skills to bear on issues directly relevant to biodefense and emerging infections, concentrating on NIAID priority pathogens. This effort is now limited by the availability of biocontainment space on the UAB campus. SEBLAB will greatly facilitate maximal national benefit from this talent, and will provide a rich environment for visiting researchers from other institutions doing research that requires biocontainment. SEBLAB's design includes flexible and secure BSL-2 and 3 laboratories, ABSL-3 housing and procedure space, and offices. In the event of a bioterrorism emergency, SEBLAB will provide surge capacity for diagnostics and other necessary analyses to enhance state and regional public health responses.
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1 |
2003 |
Marchase, Richard Banfield |
C06Activity Code Description: To provide matching Federal funds, up to 75%, for construction or major remodeling, to create new research facilities. In addition to basic research laboratories this may include, under certain circumstances, animal facilities and/or limited clinical facilities where they are an integral part of an overall research effort. |
Extramural Research Facilities Improvement Construction @ University of Alabama At Birmingham
[unreadable] DESCRIPTION (provided by applicant): [unreadable] This proposal request $4 million NCRR funding to complete 34,018 sq. ft. of animal housing, procedure, and support space. When matched with $6.25 million from the University of Alabama at Birmingham (UAB) this award will fit-out two of five floors in the Research Support Building (RSB). The RSB will serve as one of to UAB hubs for animal support, and construction is about to begin. The award will be used to finish and equip 17,009 sq. ft. of animal support space (cage wash, autoclave, receiving, necropsy, storage) on Level 1 and an equal amount of animal housing and procedure space on Level 5. Level 5 is designed at ABSL-2 but has 1, 835 sq. ft. ABSL-3/BSL3 biocontainment suite. The project will provide space for over 8, 000 small mouse cages in ventilated racks. The space will be used primarily to cage the animals of investigators in the UAB Immunology and Autoimmunity Program, a program with over $75 million in active and pending PHS funding. In addition, the project will relieve the campus-wide pressure on animal space caused by the ongoing 14 percent annual increase in the daily mouse census. It also supports four mouse core facilities servicing many of these PHS investigators (gnotobiotics, hybridoma/phage display, mapping of complex traits, and propagation of genetically engineered mice). Lastly, this project works toward our long-term goal of centralizing animal care and procedure space so as to optimize the quality of the animal program at minimal costs to investigators and research sponsors. [unreadable] [unreadable]
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1 |
2004 — 2007 |
Marchase, Richard Banfield |
R01Activity Code Description: To support a discrete, specified, circumscribed project to be performed by the named investigator(s) in an area representing his or her specific interest and competencies. |
Cytoplasmic Glycosylation and Hypovolemic Stress @ University of Alabama At Birmingham
DESCRIPTION (provided by applicant): The hypothesis to be tested here is that interventions following trauma that increase the glycosylation of nucleocytoplasmic proteins in cardiomyocytes decrease the cellular damage that would otherwise contribute to hypovolemic circulatory collapse. Our preliminary data demonstrate that in a rat model of hypovolemic stress, the infusion of glucosamine leads to a striking improvement in post-trauma function. We also demonstrate that glucosamine is protective in isolated heart models of ischemia/reperfusion and calcium overload. We propose that this protection results from an amplification of a natural, stress-activated, pro-survival pathway triggered by increased flux through the hexosamine biosynthesis pathway (HBP). The capacity to transduce an increase in the HBP's primary product, UDP-GIcNAc, into complex cellular responses comes about because O-GIcNAc transferase (OGT) is activated and recognizes distinct proteins as levels of UDP-GIcNAc increase. The identification of heat shock protein (HSP)-70 as an HBP-induced protein substrate for the OGT and the finding that HSP's associate with other proteins in part based on their ability to bind O-GIcNAc position this pathway at the center of a primary cellular response to stress. In support of this central role for O-GIcNAc in cellular protection, it is now clear that various stresses lead to increases in levels of both UDP-GIcNAc and protein-associated O-GIcNAc. We suggest that interventions that amplify and/or accelerate increases in O-GIcNAc greatly decrease the damage resulting from stress, particularly hypovolemic stress. Our Specific Aims are: to optimize recovery in the rat model of hypovolemic stress through interventions that increase UDP-GIcNAc and to test the efficacy of these interventions in swine; to test the hypothesis that the mechanism by which increased UDP-GIcNAc leads to its ameliorating effects is because of an increase in O-GIcNAc on nuclear and cytoplasmic proteins; to document and identify the nucleocytoplasmic proteins in the heart that are preferentially derivatized with O-GIcNAc when protection is optimized in the hypovolemic rat model; to investigate the decrease in calcium influx seen with increased UDP-GIcNAc in mediating improved recoveries.
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1 |
2004 |
Marchase, Richard Banfield |
C06Activity Code Description: To provide matching Federal funds, up to 75%, for construction or major remodeling, to create new research facilities. In addition to basic research laboratories this may include, under certain circumstances, animal facilities and/or limited clinical facilities where they are an integral part of an overall research effort. |
Extramural Research Facilities Improvement @ University of Alabama At Birmingham
[unreadable] DESCRIPTION (provided by applicant): [unreadable] This proposal requests $4 million of NCRR funding to assist in the completion of 45,996 gsf of research and research support space. When matched with $5.746 million from the University of Alabama at Birmingham, this award will fit out two of twelve levels (Levels 6 and 11) in the Richard C. and Annette N. Shelby Interdisciplinary Biomedical Research Building as Phase 2 of construction. The first aim is to finish and equip 22,998 gsf of space for the Mucosal Immunology Program on Level 6. This space will house an interdisciplinary research group with the relocation of six current investigators from three departments and two future recruits. The long-term objective of this group is to advance the understanding of mucosal immunology in human diseases, with a particular emphasis on inflammatory bowel disease. The second aim is to finish and equip 22,998 gsf of space for the Interdisciplinary Neurosciences Program. The goal of this unit is to advance the understanding of learning and memory, behavior, cognition, and brain cancers. Six investigators from four departments will occupy Level 11, and space will be provided for two recruits. This project will also provide space for a shared imaging facility. Space liberated by the relocation of these investigators will provide local relief for space-limited departments and an opportunity to modernize older space to create additional, efficient worksites for research. This project will thus provide state-of-the-art facilities to support federally funded research in areas emerging as high priorities for the University and the nation. [unreadable] [unreadable]
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1 |
2004 |
Marchase, Richard Banfield |
C06Activity Code Description: To provide matching Federal funds, up to 75%, for construction or major remodeling, to create new research facilities. In addition to basic research laboratories this may include, under certain circumstances, animal facilities and/or limited clinical facilities where they are an integral part of an overall research effort. |
Extramural Research Facilities Improvement: Aids @ University of Alabama At Birmingham |
1 |
2004 |
Marchase, Richard Banfield |
C06Activity Code Description: To provide matching Federal funds, up to 75%, for construction or major remodeling, to create new research facilities. In addition to basic research laboratories this may include, under certain circumstances, animal facilities and/or limited clinical facilities where they are an integral part of an overall research effort. |
Extramural Research Facilities Improvement: Environmental Health @ University of Alabama At Birmingham |
1 |
2004 |
Marchase, Richard Banfield |
C06Activity Code Description: To provide matching Federal funds, up to 75%, for construction or major remodeling, to create new research facilities. In addition to basic research laboratories this may include, under certain circumstances, animal facilities and/or limited clinical facilities where they are an integral part of an overall research effort. |
Extramural Research Facilities Improvement: Immunology @ University of Alabama At Birmingham |
1 |
2004 |
Marchase, Richard Banfield |
C06Activity Code Description: To provide matching Federal funds, up to 75%, for construction or major remodeling, to create new research facilities. In addition to basic research laboratories this may include, under certain circumstances, animal facilities and/or limited clinical facilities where they are an integral part of an overall research effort. |
Extramural Research Facilities Improvement: Neuroscience @ University of Alabama At Birmingham |
1 |
2004 |
Marchase, Richard Banfield |
C06Activity Code Description: To provide matching Federal funds, up to 75%, for construction or major remodeling, to create new research facilities. In addition to basic research laboratories this may include, under certain circumstances, animal facilities and/or limited clinical facilities where they are an integral part of an overall research effort. |
Extramural Research Facilities Improvement: Physiology @ University of Alabama At Birmingham |
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