A5032136004- Heat shock proteins research
- Prion Diseases and Protein Misfolding
- Protein Structure and Dynamics
- Enzyme Structure and Function
- Neutrophil, Myeloperoxidase and Oxidative Mechanisms
- Heme Oxygenase-1 and Carbon Monoxide
- Nitric Oxide and Endothelin Effects
- Genetics, Aging, and Longevity in Model Organisms
- Hemoglobin structure and function
- Proteins in Food Systems
- Endoplasmic Reticulum Stress and Disease
- Alzheimer's disease research and treatments
- Metabolism and Genetic Disorders
- Fungal and yeast genetics research
- Biofuel production and bioconversion
- Iron Metabolism and Disorders
- Cell Adhesion Molecules Research
- Protein Hydrolysis and Bioactive Peptides
- Mitochondrial Function and Pathology
- Erythrocyte Function and Pathophysiology
- High Altitude and Hypoxia
- Nitrogen and Sulfur Effects on Brassica
- ATP Synthase and ATPases Research
- Neuroinflammation and Neurodegeneration Mechanisms
- Plant Genetic and Mutation Studies
Medical College of Wisconsin
2022-2024
Cleveland Clinic Lerner College of Medicine
2018-2021
University of Pennsylvania
2008-2019
Massachusetts Institute of Technology
2017
University of Chicago
2008
Untangling aggregates one step at a time Conserved AAA+ protein complexes exploit adenosine triphosphate hydrolysis to unfold and disaggregate their substrates in response cell stress, but exactly how they do this has been unclear. Gates et al. determined high-resolution cryo-electron microscopy structures of the Hsp104 disaggregase bound an unfolded polypeptide substrate its channel. The reveal interactions two different translocation states. undergoes conformational changes that drive...
Transferring the prokaryotic enzyme nitrogenase into a eukaryotic host with final aim of developing N2 fixing cereal crops would revolutionize agricultural systems worldwide. Targeting it to mitochondria has potential advantages because organelle's high O2 consumption and presence bacterial-type iron-sulfur cluster biosynthetic machinery. In this study, we constructed 96 strains Saccharomyces cerevisiae in which transcriptional units comprising nine Azotobacter vinelandii nif genes...
Soluble guanylyl cyclase (sGC) is a key component of NO–cGMP signaling in mammals. Although heme must bind the sGC β1 subunit (sGCβ) for to function, how delivered sGCβ remains unknown. Given that GAPDH displays properties chaperone inducible NO synthase, here we investigated whether delivery apo-sGCβ involves GAPDH. We utilized an reporter construct, tetra-Cys sGCβ, whose insertion can be followed by fluorescence quenching live cells, assessed lowering cell expression impacts delivery, and...
Maturation of adult (α2β2) and fetal hemoglobin (α2γ2) tetramers requires that heme be incorporated into each globin. While alpha (Hb-α) relies on a specific erythroid chaperone (alpha Hb-stabilizing protein, AHSP), the other chaperones may help mature partner globins (Hb-γ or Hb-β) in cells, enable nonerythroid cells to express Hb, are unknown. We investigated role for heat-shock protein 90 (hsp90) Hb maturation precursor naturally Hb-α with either Hb-γ (K562 HiDEP-1 cells) Hb-β (HUDEP-2)...
Hsp104 is an AAA+ protein disaggregase, which can be potentiated via diverse mutations in its autoregulatory middle domain (MD) to mitigate toxic misfolding of TDP-43, FUS, and α-synuclein implicated fatal neurodegenerative disorders. Problematically, MD variants exhibit off-target toxicity. Here, we mine disaggregase sequence space safely enhance activity single nucleotide-binding 1 (NBD1) or NBD2. Like variants, NBD counter toxicity elevated ATPase activity. Unlike non-toxic NBD1 NBD2...
GAPDH, a heme chaperone, has been previously implicated in the incorporation of into iNOS and soluble guanylyl cyclase (sGC). Since sGC is critical for myoglobin (Mb) heme-maturation, we investigated role GAPDH maturation this globin, as well hemoglobins α, β, γ. Utilizing cell culture systems, found that overexpression wild-type increased, whereas mutants H53A K227A decreased, content Mb Hbα Hbβ. Overexpression fully recovered heme-maturation inhibition observed with mutants. Partial rescue...
Hsp104 is a hexameric AAA
Hsp104 is a hexameric AAA+ protein1 from yeast, which couples ATP hydrolysis to protein disaggregation2-10 (Fig. 1). This activity imparts two key selective advantages. First, renaturation of disordered aggregates by empowers yeast survival after various protein-misfolding stresses, including heat shock3,5,11,12. Second, remodeling cross-beta amyloid fibrils enables exploit myriad prions (infectious amyloids) as reservoir beneficial and heritable phenotypic variation13-22. Remarkably,...
Recent Hsp104 structural studies have reported both planar and helical models of the hexameric structure. The conformation monomers within hexamer is affected by nucleotide ligation. After nucleotide-driven formation, Hsp104-catalyzed disruption protein aggregates requires binding to peptide substrate. Here, we examine oligomeric state its competency in absence presence ADP, ATPγS, AMPPNP, or AMPPCP. Surprisingly, found that only ATPγS facilitates avid Hsp104. We propose modulation between...
NADPH oxidase 5 (NOX5) is a transmembrane signaling enzyme that produces superoxide in response to elevated cytosolic calcium. In addition its association with numerous human diseases, NOX5 has recently been discovered play crucial roles the immune and cardiovascular system. Details of maturation, specifically changes intracellular heme levels have remained unclear. Here we establish an experimental system mammalian cells allows us probe influence availability on ROS production by NOX5. We...
NADPH oxidase 5 (NOX5) is a transmembrane oxidative signaling enzyme which produces superoxide in response to intracellular calcium flux. Increasing evidence indicates that NOX5 involved variety of physiological processes as well human disease, however, details pathways and targets mediated modifications remain poorly resolved. Actin dynamics have previously been shown be modulated by modification, direct connection expression activity has not fully explored. Here we show actin interact the...
Hsp104 is a hexameric AAA+ protein1 from yeast, which couples ATP hydrolysis to protein disaggregation2-10 (Fig. 1). This activity imparts two key selective advantages. First, renaturation of disordered aggregates by empowers yeast survival after various protein-misfolding stresses, including heat shock3,5,11,12. Second, remodeling cross-beta amyloid fibrils enables exploit myriad prions (infectious amyloids) as reservoir beneficial and heritable phenotypic variation13-22. Remarkably,...
<title>Abstract</title> Heme is an iron-containing cofactor essential for life. In eukaryotes heme generated in the mitochondria and must leave this organelle to reach protein targets other cell compartments. Mitochondrial binding by cytosolic GAPDH was recently found distribution eukaryotic cells. Here, we sought uncover how mitochondrial reaches GAPDH. Experiments involving a human line novel reporter construct whose live cells can be followed fluorescence revealed that transmembrane...
Abstract Heme is an iron-containing cofactor essential for life. In eukaryotes heme generated in the mitochondria and must leave this organelle to reach protein targets other cell compartments. Mitochondrial binding by cytosolic GAPDH was recently found distribution eukaryotic cells. Here, we sought uncover how mitochondrial reaches GAPDH. Experiments involving a human line novel reporter construct whose live cells can be followed fluorescence revealed that transmembrane FLVCR1b exclusively...