A5059113011- Amyotrophic Lateral Sclerosis Research
- Neurogenetic and Muscular Disorders Research
- Ion channel regulation and function
- Muscle Physiology and Disorders
- Neurological diseases and metabolism
- Cardiac electrophysiology and arrhythmias
- Parkinson's Disease Mechanisms and Treatments
- Neuroscience and Neural Engineering
- Genetic Neurodegenerative Diseases
- biodegradable polymer synthesis and properties
- Trace Elements in Health
- Prion Diseases and Protein Misfolding
- RNA Research and Splicing
- Cardiomyopathy and Myosin Studies
- Alzheimer's disease research and treatments
- Cholinesterase and Neurodegenerative Diseases
- Neuroscience and Neuropharmacology Research
- Cancer-related gene regulation
- Ion Transport and Channel Regulation
- TGF-β signaling in diseases
- Pluripotent Stem Cells Research
- CAR-T cell therapy research
- CRISPR and Genetic Engineering
- Lanthanide and Transition Metal Complexes
- Mitochondrial Function and Pathology
University of Massachusetts Chan Medical School
2014-2024
Neurology, Inc
2005
San Jose State University
2005
University of California, Los Angeles
2005
Daresbury Laboratory
2004
The University of Melbourne
2002
Royal Women's Hospital
2002
CFD Research Corporation (United States)
2002
Harvard University
1995-1999
Massachusetts General Hospital
1996-1999
Mutations in the RNA-binding protein FUS (fused sarcoma) are linked to amyotrophic lateral sclerosis (ALS), but mechanism by which these mutants cause motor neuron degeneration is not known. We report a novel ALS truncation mutant (R495X) that leads relatively severe clinical phenotype compared with missense mutations. Expression of R495X FUS, abrogates putative nuclear localization signal at C-terminus HEK-293 cells and zebrafish spinal cord caused striking cytoplasmic accumulation greater...
Over 90 different mutations in the gene encoding copper/zinc superoxide dismutase (SOD1) cause ∼2% of amyotrophic lateral sclerosis (ALS) cases by an unknown mechanism. We engineered 14 human ALS-related SOD1 mutants and obtained high yields biologically metallated proteins from Sf21 insect cell expression system. Both wild type mutant "as isolated" variants were deficient copper heterogeneous native gel electrophoresis. By contrast, although three SOD1s with substitutions near metal binding...
We report the thermal stability of wild type (WT) and 14 different variants human copper/zinc superoxide dismutase (SOD1) associated with familial amyotrophic lateral sclerosis (FALS). Multiple endothermic unfolding transitions were observed by differential scanning calorimetry for partially metallated SOD1 enzymes isolated from a baculovirus system. correlated metal ion contents occurrence distinct melting transitions. Altered upon reduction copper dithionite identified resulting...
More than 90 point mutations in human CuZn superoxide dismutase lead to the development of familial amyotrophic lateral sclerosis, known also as motor neuron disease. A growing body evidence suggests that a subset located close dimeric interface can major destabilization mutant enzymes. We have determined crystal structures Ala4Val (A4V) and Ile113Thr (I113T) mutants 1.9 1.6 Å, respectively. In A4V structure, small changes at dimer result substantial reorientation two monomers. This effect...
FUS/TLS is a nucleic acid binding protein that, when mutated, can cause subset of familial amyotrophic lateral sclerosis (fALS). Although normally located predominantly in the nucleus, pathogenic mutant forms traffic to, and form inclusions in, cytoplasm affected spinal motor neurons or glia. Here we report yeast model human expression that recapitulates multiple salient features pathology disease-causing proteins, including nuclear to cytoplasmic translocation, inclusion formation,...
Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disorder characterized by progressive motor neuron death. More than 90 mutations in the copper-zinc superoxide dismutase (SOD1) gene cause subset of familial ALS. Toxic properties have been proposed for ALS-linked SOD1 mutants, but nature toxicity has not clearly specified. Cytoplasmic inclusion bodies containing mutant and number other proteins are pathological hallmark SOD1-mediated ALS, whether such aggregates toxic to neurons...
Embryonic muscle development permits the study of contractile protein gene regulation during cellular differentiation. To distinguish appearance particular actin mRNAs chicken myogenesis, we have constructed DNA probes from transcribed 3' noncoding region single-copy alpha-skeletal, alpha-cardiac, and beta-cytoplasmic genes. Hybridization experiments showed that at day 10 in ovo (stage 36), embryonic hindlimbs contain low levels mRNA, predominantly consisting alpha-cardiac beta-actin...
Dying-back degeneration of motor neuron axons represents an established feature familial amyotrophic lateral sclerosis (FALS) associated with superoxide dismutase 1 (SOD1) mutations, but axon-autonomous effects pathogenic SOD1 remained undefined. Characteristics neurons affected in FALS include abnormal kinase activation, aberrant neurofilament phosphorylation, and fast axonal transport (FAT) deficits, functional relationships among these events were unclear. Experiments isolated squid...
Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease. Mutations in Cu/Zn superoxide dismutase (SOD1) are responsible for approximately 20 % of the familial ALS cases. ALS-causing SOD1 mutants display gain-of-toxicity phenotype, but nature this toxicity still not fully understood. The Ras GTPase-activating protein-binding protein G3BP1 plays critical role stress granule dynamics. Alterations dynamics granules have been reported several other forms unrelated to SOD1. To our...
The relative stabilities and structural properties of a representative set 20 ALS-mutant Cu,Zn-superoxide dismutase apoproteins were examined by using differential scanning calorimetry hydrogen-deuterium (H/D) exchange followed MS. Contrary to recent reports from other laboratories, we found that are not universally destabilized the disease-causing mutations. For example, several with substitutions at or near metal binding region (MBR) (MBR mutants) exhibited melting temperatures (Tm) in...
Missense mutations in the skeletal muscle Na+ channel alpha subunit occur several heritable forms of myotonia and periodic paralysis. Distinct phenotypes arise from at two sites within III-IV cytoplasmic loop: without weakness due to substitutions glycine 1306, plus caused by a mutation threonine 1313. Heterologous expression HEK cells showed that either site disrupted inactivation, as reflected slower inactivation rates, shifts steady-state larger persistent currents. For T1313M, however,...
Mutations in the gene encoding human copper-zinc superoxide dismutase (SOD1) cause a dominant form of progressive neurodegenerative disease amyotrophic lateral sclerosis. Transgenic mice expressing G85R SOD1 variant develop paralytic symptoms concomitant with appearance SOD1-enriched proteinaceous inclusions their neural tissues. The process(es) through which misfolding or aggregation induces motor neuron toxicity is not understood. Here we present structures determined by single crystal...
More than 100 different mutations in the gene encoding Cu,Zn-superoxide dismutase (SOD1) cause preferential motor neuron degeneration familial amyotrophic lateral sclerosis (ALS). Although cellular target(s) of mutant SOD1 toxicity have not been precisely specified, evidence to date supports hypothesis that ALS-related may increase burden partially unfolded species. Influences destabilize vivo include impaired metal ion binding, reduction intrasubunit disulfide bond, or oxidative...
Top-down and bottom-up mass spectrometry methods can generate gas phase fragments use these to identify proteins. methods, in addition, provide the of protein itself therefore additional structural information. Despite conceptual advantage top-down market share belongs as a result their more robust sample preparation, fragmentation, data processing methods. Here we report improved fragmentation for spectrometry. Specifically funnel-skimmer dissociation, variation nozzle-skimmer compare its...
The mechanisms by which mutant variants of Cu/Zn-superoxide dismutase (SOD1) cause familial amyotrophic lateral sclerosis are not clearly understood. Evidence to date suggests that altered conformations SOD1s trigger perturbations cellular homeostasis ultimately motor neuron degeneration. In this study we correlated the metal contents and disulfide bond status purified wild-type (WT) SOD1 proteins changes in electrophoretic mobility surface hydrophobicity as detected...
At least 119 mutations in the gene encoding copper/zinc superoxide dismutase (SOD1) cause amyotrophic lateral sclerosis by an unidentified toxic gain of function. We compared dynamic properties 13 as-isolated, partially metallated, SOD1 variant enzymes using hydrogen-deuterium exchange. identified a shared property these familial sclerosis-related variants, namely structural and change affecting electrostatic loop (loop VII) SOD1. Furthermore, variants that have severely compromised metal...