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Papers by Kathleen Boesze-Battaglia
Journal of Experimental Biology, 1997
Photoreceptor rod cells and blood platelets are remarkably different, yet both illustrate a simil... more Photoreceptor rod cells and blood platelets are remarkably different, yet both illustrate a similar phenomenon. Both are strongly affected by membrane cholesterol, and the distribution of cholesterol in the membranes of both cell types is determined by the lipid composition within the membranes. In rod cells, cholesterol strongly inhibits rhodopsin activity. The relatively higher level of cholesterol in the plasma membrane serves to inhibit, and thereby conserve, the activity of rhodopsin, which becomes fully active in the low-cholesterol environment of the disk membranes of these same cells. This physiologically important partitioning of cholesterol between disk membranes and plasma membranes occurs because the disk membranes are enriched with phosphatidylethanolamine, thus providing a thermodynamically unfavorable environment for the sterol. Cholesterol enrichment of platelets renders these cells more responsive to stimuli of aggregation. Stimuli for platelet aggregation cause a r...
Cellular Physiology and Biochemistry, 2020
This article is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 I... more This article is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License (CC BY-NC-ND). Usage and distribution for commercial purposes as well as any distribution of modified material requires written permission.
Objectives: Nitrogen-containing bisphosphonates (nBPs) are commonly used to manage skeletal event... more Objectives: Nitrogen-containing bisphosphonates (nBPs) are commonly used to manage skeletal events of cancer metastasis because internalization of nBP by osteoclasts results in apoptosis. nBP therapy is often complicated by osteonecrosis of the jaws (ONJ), but the underlying pathophysiology is yet to be fully elucidated. Bone mesenchymal stem cells (MSCs) are postnatal stem cells that can form multiple cell types including osteoblasts needed to maintain osteoclast-osteoblast balance during bone remodeling. If the jaw is disparately targeted in ONJ, disproportionate nBP uptake by orofacial/jaw MSCs (OFMSCs) with consequent apoptosis will reduce the ‘pool’ of osteoprogenitors essential for bone healing and homeostasis. We hypothesized that internalized nBP is processed differentially by OFMSCs relative to a non-oral skeletal site. Methods: Sub-confluent normal human MSCs from the jaw (OFMSCs) and iliac crest (ICMSCs) cultured at 104 cells/cm2 were exposed to 0.25mg/ml fluorescently la...
Infection and Immunity, 2015
Induction of cell cycle arrest in lymphocytes following exposure to the Aggregatibacter actinomyc... more Induction of cell cycle arrest in lymphocytes following exposure to the Aggregatibacter actinomycetemcomitans cytolethal distending toxin (Cdt) is dependent upon the integrity of lipid membrane microdomains. Moreover, we have previously demonstrated that the association of Cdt with target cells involves the CdtC subunit which binds to cholesterol via a cholesterol recognition amino acid consensus sequence (CRAC site). In this study, we demonstrate that the active Cdt subunit, CdtB, also is capable of binding to large unilamellar vesicles (LUVs) containing cholesterol. Furthermore, CdtB binding to cholesterol involves a similar CRAC site as that demonstrated for CdtC. Mutation of the CRAC site reduces binding to model membranes as well as toxin binding and CdtB internalization in both Jurkat cells and human macrophages. A concomitant reduction in Cdt-induced toxicity was also noted, indicated by reduced cell cycle arrest and apoptosis in Jurkat cells and a reduction in the proinflamm...
Infection and Immunity, 2015
The cytolethal distending toxin (Cdt) is produced from a number of bacteria capable of causing in... more The cytolethal distending toxin (Cdt) is produced from a number of bacteria capable of causing infection and inflammatory disease. Our previous studies with Actinobacillus actinomycetemcomitans Cdt demonstrate not only that the active toxin subunit functions as a phosphatidylinositol-3,4,5-triphosphate (PIP3) phosphatase but also that macrophages exposed to the toxin were stimulated to produce proinflammatory cytokines. We now demonstrate that the Cdt-induced proinflammatory response involves the activation of the NLRP3 inflammasome. Specific inhibitors and short hairpin RNA (shRNA) were employed to demonstrate requirements for NLRP3 and ASC as well as caspase-1. Furthermore, Cdt-mediated inflammasome activation is dependent upon upstream signals, including reactive oxygen species (ROS) generation and Cdt-induced increases in extracellular ATP levels. Increases in extracellular ATP levels contribute to the activation of the P2X 7 purinergic receptor, leading to K + efflux. The relat...
Experimental Eye Research, 2002
Peripherin/rds is proposed to function as a fusion protein within the rod outer segment and a fus... more Peripherin/rds is proposed to function as a fusion protein within the rod outer segment and a fusion domain has been mapped to amino acids 311-325 within the C-terminus. To map regions within peripherin/rds required for membrane fusion a series of C-terminal mutants was analyzed. Madin Darby canine kidney cells were transiently transfected with an Xpress or FLAG epitope tagged peripherin/rds (wt) and three mutants of peripherin/rds. The mutants selected were a P296T mutant (replacement of the proline at position 296 with a threonine) and two C-terminal deletion mutants (one lacking the terminal 10 amino acids, Δ10 and one lacking the terminal 50 amino acids, Δ50). The wt protein, the P296T and Δ10 mutants were detected on SDS-PAGE as 84 kDa dimers, that resolved into 38-42 kDa monomers under reducing conditions. The Δ50 mutant
Experimental Eye Research, 1989
Current Pharmaceutical Design, 2011
Biophysical Journal, 2010
Biochemistry, 2007
The C-terminus of the intracellular retinal rod outer segment disk protein peripherin-2 binds to ... more The C-terminus of the intracellular retinal rod outer segment disk protein peripherin-2 binds to membranes, adopts a helical conformation, and promotes membrane fusion, which suggests an analogy to the structure and function of viral envelope fusion proteins. Nuclear magnetic resonance (NMR) data and fluorescence data show that a 63-residue polypeptide comprising the C-terminus of bovine peripherin-2 (R284-G346) binds to the membrane mimetic, dodecylphosphocholine micelles. High-resolution NMR studies reveal that although this C-terminal fragment is unstructured in solution, the same fragment adopts helical structure when bound to the micelles. The C-terminus may be a member of the class of intrinsically unstructured protein domains. Using methods developed for the G-protein coupled receptor rhodopsin, a model for the structure of the transmembrane domain of peripherin-2 was constructed. Previously published data showed that both peripherin-2 and viral fusion proteins are transmembrane proteins that promote membrane fusion and have a fusion peptide sequence within the protein that independently promotes membrane fusion. Furthermore, the fusion-active sequence of peripherin-2 exhibits a sequence motif that matches the viral fusion peptide of influenza hemagglutinin (HA). These observations collectively suggest that the mechanism of intracellular membrane fusion induced by peripherin-2 and the mechanism of enveloped viral fusion may have features in common.
Molecular Neurobiology, 2014
A main requisite in the phagocytosis of ingested material is a coordinated series of maturation s... more A main requisite in the phagocytosis of ingested material is a coordinated series of maturation steps which lead to the degradation of ingested cargo.
Molecular Oral Microbiology, 2015
Porphyromonas gingivalis often subverts host cell autophagic processes for its own survival. Our ... more Porphyromonas gingivalis often subverts host cell autophagic processes for its own survival. Our previous studies document the association of the cargo sorting protein, melanoregulin (MREG), with its binding partner, the autophagic protein, microtubule-associated protein 1 light chain 3 (LC3) in macrophages incubated with P. gingivalis (strain 33277). Differences in the lipid A moiety of lipopolysaccharide (LPS) affects the virulence of P. gingivalis; penta-acylated LPS1690 , is a weak TLR4 agonist compared to E. coli LPS, whereas tetra-acylated LPS1435/1449 acts as an LPS1690 antagonist. To determine how P. gingivalis LPS1690 affects autophagy we assessed LC3- and MREG-dependent processes in green fluorescent protein (GFP)-LC3 expressing Saos-2 cells. LPS1690 stimulated the formation of very large LC3-positive vacuoles and MREG puncta. This LPS1690 -mediated LC3 lipidation decreased in the presence of LPS1435/1449 . When Saos-2 cells were incubated with P. gingivalis the bacteria internalized but did not traffic to GFP-LC3-positive structures. Nevertheless, increases in LC3 lipidation and MREG puncta were observed. Collectively, these results suggest that P. gingivalis internalization is not necessary for LC3 lipidation. Primary human gingival epithelial cells isolated from periodontitis patients incubated with P. gingivalis showed both LC3II and MREG puncta while cells from disease-free individuals exhibited little colocalization of these two proteins. These results suggest that the prevalence of a particular LPS moiety may modulate the degradative capacity of host cells thus influencing bacterial survival. This article is protected by copyright. All rights reserved.
Journal of Biological Chemistry, 2017
Communications Biology
Late-onset retinal degeneration (L-ORD) is an autosomal dominant disorder caused by a missense su... more Late-onset retinal degeneration (L-ORD) is an autosomal dominant disorder caused by a missense substitution in CTRP5. Distinctive clinical features include sub-retinal pigment epithelium (RPE) deposits, choroidal neovascularization, and RPE atrophy. In induced pluripotent stem cells-derived RPE from L-ORD patients (L-ORD-iRPE), we show that the dominant pathogenic CTRP5 variant leads to reduced CTRP5 secretion. In silico modeling suggests lower binding of mutant CTRP5 to adiponectin receptor 1 (ADIPOR1). Downstream of ADIPOR1 sustained activation of AMPK renders it insensitive to changes in AMP/ATP ratio resulting in defective lipid metabolism, reduced Neuroprotectin D1(NPD1) secretion, lower mitochondrial respiration, and reduced ATP production. These metabolic defects result in accumulation of sub-RPE deposits and leave L-ORD-iRPE susceptible to dedifferentiation. Gene augmentation of L-ORD-iRPE with WT CTRP5 or modulation of AMPK, by metformin, re-sensitize L-ORD-iRPE to changes ...
Investigative Ophthalmology & Visual Science, 2002
Investigative Ophthalmology & Visual Science, 2016
Journal of Experimental Biology, 1997
Photoreceptor rod cells and blood platelets are remarkably different, yet both illustrate a simil... more Photoreceptor rod cells and blood platelets are remarkably different, yet both illustrate a similar phenomenon. Both are strongly affected by membrane cholesterol, and the distribution of cholesterol in the membranes of both cell types is determined by the lipid composition within the membranes. In rod cells, cholesterol strongly inhibits rhodopsin activity. The relatively higher level of cholesterol in the plasma membrane serves to inhibit, and thereby conserve, the activity of rhodopsin, which becomes fully active in the low-cholesterol environment of the disk membranes of these same cells. This physiologically important partitioning of cholesterol between disk membranes and plasma membranes occurs because the disk membranes are enriched with phosphatidylethanolamine, thus providing a thermodynamically unfavorable environment for the sterol. Cholesterol enrichment of platelets renders these cells more responsive to stimuli of aggregation. Stimuli for platelet aggregation cause a r...
Cellular Physiology and Biochemistry, 2020
This article is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 I... more This article is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License (CC BY-NC-ND). Usage and distribution for commercial purposes as well as any distribution of modified material requires written permission.
Objectives: Nitrogen-containing bisphosphonates (nBPs) are commonly used to manage skeletal event... more Objectives: Nitrogen-containing bisphosphonates (nBPs) are commonly used to manage skeletal events of cancer metastasis because internalization of nBP by osteoclasts results in apoptosis. nBP therapy is often complicated by osteonecrosis of the jaws (ONJ), but the underlying pathophysiology is yet to be fully elucidated. Bone mesenchymal stem cells (MSCs) are postnatal stem cells that can form multiple cell types including osteoblasts needed to maintain osteoclast-osteoblast balance during bone remodeling. If the jaw is disparately targeted in ONJ, disproportionate nBP uptake by orofacial/jaw MSCs (OFMSCs) with consequent apoptosis will reduce the ‘pool’ of osteoprogenitors essential for bone healing and homeostasis. We hypothesized that internalized nBP is processed differentially by OFMSCs relative to a non-oral skeletal site. Methods: Sub-confluent normal human MSCs from the jaw (OFMSCs) and iliac crest (ICMSCs) cultured at 104 cells/cm2 were exposed to 0.25mg/ml fluorescently la...
Infection and Immunity, 2015
Induction of cell cycle arrest in lymphocytes following exposure to the Aggregatibacter actinomyc... more Induction of cell cycle arrest in lymphocytes following exposure to the Aggregatibacter actinomycetemcomitans cytolethal distending toxin (Cdt) is dependent upon the integrity of lipid membrane microdomains. Moreover, we have previously demonstrated that the association of Cdt with target cells involves the CdtC subunit which binds to cholesterol via a cholesterol recognition amino acid consensus sequence (CRAC site). In this study, we demonstrate that the active Cdt subunit, CdtB, also is capable of binding to large unilamellar vesicles (LUVs) containing cholesterol. Furthermore, CdtB binding to cholesterol involves a similar CRAC site as that demonstrated for CdtC. Mutation of the CRAC site reduces binding to model membranes as well as toxin binding and CdtB internalization in both Jurkat cells and human macrophages. A concomitant reduction in Cdt-induced toxicity was also noted, indicated by reduced cell cycle arrest and apoptosis in Jurkat cells and a reduction in the proinflamm...
Infection and Immunity, 2015
The cytolethal distending toxin (Cdt) is produced from a number of bacteria capable of causing in... more The cytolethal distending toxin (Cdt) is produced from a number of bacteria capable of causing infection and inflammatory disease. Our previous studies with Actinobacillus actinomycetemcomitans Cdt demonstrate not only that the active toxin subunit functions as a phosphatidylinositol-3,4,5-triphosphate (PIP3) phosphatase but also that macrophages exposed to the toxin were stimulated to produce proinflammatory cytokines. We now demonstrate that the Cdt-induced proinflammatory response involves the activation of the NLRP3 inflammasome. Specific inhibitors and short hairpin RNA (shRNA) were employed to demonstrate requirements for NLRP3 and ASC as well as caspase-1. Furthermore, Cdt-mediated inflammasome activation is dependent upon upstream signals, including reactive oxygen species (ROS) generation and Cdt-induced increases in extracellular ATP levels. Increases in extracellular ATP levels contribute to the activation of the P2X 7 purinergic receptor, leading to K + efflux. The relat...
Experimental Eye Research, 2002
Peripherin/rds is proposed to function as a fusion protein within the rod outer segment and a fus... more Peripherin/rds is proposed to function as a fusion protein within the rod outer segment and a fusion domain has been mapped to amino acids 311-325 within the C-terminus. To map regions within peripherin/rds required for membrane fusion a series of C-terminal mutants was analyzed. Madin Darby canine kidney cells were transiently transfected with an Xpress or FLAG epitope tagged peripherin/rds (wt) and three mutants of peripherin/rds. The mutants selected were a P296T mutant (replacement of the proline at position 296 with a threonine) and two C-terminal deletion mutants (one lacking the terminal 10 amino acids, Δ10 and one lacking the terminal 50 amino acids, Δ50). The wt protein, the P296T and Δ10 mutants were detected on SDS-PAGE as 84 kDa dimers, that resolved into 38-42 kDa monomers under reducing conditions. The Δ50 mutant
Experimental Eye Research, 1989
Current Pharmaceutical Design, 2011
Biophysical Journal, 2010
Biochemistry, 2007
The C-terminus of the intracellular retinal rod outer segment disk protein peripherin-2 binds to ... more The C-terminus of the intracellular retinal rod outer segment disk protein peripherin-2 binds to membranes, adopts a helical conformation, and promotes membrane fusion, which suggests an analogy to the structure and function of viral envelope fusion proteins. Nuclear magnetic resonance (NMR) data and fluorescence data show that a 63-residue polypeptide comprising the C-terminus of bovine peripherin-2 (R284-G346) binds to the membrane mimetic, dodecylphosphocholine micelles. High-resolution NMR studies reveal that although this C-terminal fragment is unstructured in solution, the same fragment adopts helical structure when bound to the micelles. The C-terminus may be a member of the class of intrinsically unstructured protein domains. Using methods developed for the G-protein coupled receptor rhodopsin, a model for the structure of the transmembrane domain of peripherin-2 was constructed. Previously published data showed that both peripherin-2 and viral fusion proteins are transmembrane proteins that promote membrane fusion and have a fusion peptide sequence within the protein that independently promotes membrane fusion. Furthermore, the fusion-active sequence of peripherin-2 exhibits a sequence motif that matches the viral fusion peptide of influenza hemagglutinin (HA). These observations collectively suggest that the mechanism of intracellular membrane fusion induced by peripherin-2 and the mechanism of enveloped viral fusion may have features in common.
Molecular Neurobiology, 2014
A main requisite in the phagocytosis of ingested material is a coordinated series of maturation s... more A main requisite in the phagocytosis of ingested material is a coordinated series of maturation steps which lead to the degradation of ingested cargo.
Molecular Oral Microbiology, 2015
Porphyromonas gingivalis often subverts host cell autophagic processes for its own survival. Our ... more Porphyromonas gingivalis often subverts host cell autophagic processes for its own survival. Our previous studies document the association of the cargo sorting protein, melanoregulin (MREG), with its binding partner, the autophagic protein, microtubule-associated protein 1 light chain 3 (LC3) in macrophages incubated with P. gingivalis (strain 33277). Differences in the lipid A moiety of lipopolysaccharide (LPS) affects the virulence of P. gingivalis; penta-acylated LPS1690 , is a weak TLR4 agonist compared to E. coli LPS, whereas tetra-acylated LPS1435/1449 acts as an LPS1690 antagonist. To determine how P. gingivalis LPS1690 affects autophagy we assessed LC3- and MREG-dependent processes in green fluorescent protein (GFP)-LC3 expressing Saos-2 cells. LPS1690 stimulated the formation of very large LC3-positive vacuoles and MREG puncta. This LPS1690 -mediated LC3 lipidation decreased in the presence of LPS1435/1449 . When Saos-2 cells were incubated with P. gingivalis the bacteria internalized but did not traffic to GFP-LC3-positive structures. Nevertheless, increases in LC3 lipidation and MREG puncta were observed. Collectively, these results suggest that P. gingivalis internalization is not necessary for LC3 lipidation. Primary human gingival epithelial cells isolated from periodontitis patients incubated with P. gingivalis showed both LC3II and MREG puncta while cells from disease-free individuals exhibited little colocalization of these two proteins. These results suggest that the prevalence of a particular LPS moiety may modulate the degradative capacity of host cells thus influencing bacterial survival. This article is protected by copyright. All rights reserved.
Journal of Biological Chemistry, 2017
Communications Biology
Late-onset retinal degeneration (L-ORD) is an autosomal dominant disorder caused by a missense su... more Late-onset retinal degeneration (L-ORD) is an autosomal dominant disorder caused by a missense substitution in CTRP5. Distinctive clinical features include sub-retinal pigment epithelium (RPE) deposits, choroidal neovascularization, and RPE atrophy. In induced pluripotent stem cells-derived RPE from L-ORD patients (L-ORD-iRPE), we show that the dominant pathogenic CTRP5 variant leads to reduced CTRP5 secretion. In silico modeling suggests lower binding of mutant CTRP5 to adiponectin receptor 1 (ADIPOR1). Downstream of ADIPOR1 sustained activation of AMPK renders it insensitive to changes in AMP/ATP ratio resulting in defective lipid metabolism, reduced Neuroprotectin D1(NPD1) secretion, lower mitochondrial respiration, and reduced ATP production. These metabolic defects result in accumulation of sub-RPE deposits and leave L-ORD-iRPE susceptible to dedifferentiation. Gene augmentation of L-ORD-iRPE with WT CTRP5 or modulation of AMPK, by metformin, re-sensitize L-ORD-iRPE to changes ...
Investigative Ophthalmology & Visual Science, 2002
Investigative Ophthalmology & Visual Science, 2016