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Harry R. Allcock - One of the best experts on this subject based on the ideXlab platform.

  • miscibility of choline substituted polyphosphazenes with plga and osteoblast activity on resulting blends
    Biomaterials, 2010
    Co-Authors: Arlin L. Weikel, Lakshmi S. Nair, Cato T. Laurencin, Steven G Owens, Nicole L Morozowich, Meng Deng, Harry R. Allcock
    Abstract:

    The preparation of phosphazene tissue engineering scaffolds with bioactive side groups has been accomplished using the biological buffer, choline chloride. Mixed-substituent phosphazene cyclic trimers (as model systems) and polymers with choline chloride and Glycine Ethyl Ester, alanine Ethyl Ester, valine Ethyl Ester, or phenylalanine Ethyl Ester were synthesized. Two different synthetic protocols were examined. A sodium hydride mediated route resulted in polyphosphazenes with a low choline content, while a cesium carbonate mediated process produced polyphosphazenes with higher choline content. The phosphazene structures and physical properties were studied using multinuclear NMR, differential scanning calorimetry (DSC), and gel permeation chromatography (GPC) techniques. The resultant polymers were then blended with PLGA (50:50) or PLGA (85:15) and characterized by DSC analysis and scanning electron microscopy (SEM). Polymer products obtained via the sodium hydride route produced miscible blends with both ratios of PLGA, while the cesium carbonate route yielded products with reduced blend miscibility. Heterophase hydrolysis experiments in aqueous media revealed that the polymer blends hydrolyzed to near-neutral pH media (∼5.8 to 6.8). The effect of different molecular structures on cellular adhesion showed osteoblast proliferation with an elevated osteoblast phenotype expression compared to PLGA over a 21-day culture period.

  • hydrogen bonding in blends of polyEsters with dipeptide containing polyphosphazenes
    Journal of Applied Polymer Science, 2010
    Co-Authors: Nick R. Krogman, Lakshmi S. Nair, Cato T. Laurencin, Arlin L. Weikel, Katherine A. Kristhart, Syam P. Nukavarapu, Nhu Q Nguyen, Harry R. Allcock
    Abstract:

    New biomedically erodible polymer composites were investigated. Polyphosphazenes containing the dipeptide side groups alanyl–Glycine Ethyl Ester, valinyl–Glycine Ethyl Ester, and phenylalanyl–Glycine Ethyl Ester were blended with poly(lactide-co-glycolide) (PLGA) with lactic to glycolic acid ratios of 50 : 50 [PLGA (50 : 50)] and 85 : 15 [PLGA (85 : 15)] with solution-phase techniques. Each dipeptide Ethyl Ester side group contains two NH protons that are capable of hydrogen bonding with the carbonyl functions of PLGA. Polyphosphazenes that contain only the dipeptide Ethyl Ester groups are insoluble in organic solvents and are thus unsuitable for solution-phase composite formation. To ensure solubility during and after synthesis, cosubstituted polymers with both dipeptide Ethyl Ester and Glycine or alanine Ethyl Ester side groups were used. Solution casting or electrospinning was used to fabricate polymer blend matrices with different ratios of polyphosphazene to polyEster, and their miscibilities were estimated with differential scanning calorimetry and scanning electron microscopy techniques. Polyphosphazenes with alanyl–Glycine Ethyl Ester side groups plus the second cosubstituent were completely miscible with PLGA (50 : 50) and PLGA (85 : 15) when processed via solution-casting techniques. This suggests that the hydrogen-bonding protons in alanyl–Glycine Ethyl Ester have access to the oxygen atoms of the carbonyl units in PLGA. However, when the same pair of polymers was electrospun from solution, the polymers proved to be immiscible. Solution-cast miscible polymer blends were obtained from PLGA (50 : 50) plus the polyphosphazene that was cosubstituted with valinyl–Glycine Ethyl Ester and Glycine Ethyl Ester side groups. © 2009 Wiley Periodicals, Inc. J Appl Polym Sci, 2010

  • polyphosphazenes that contain dipeptide side groups synthesis characterization and sensitivity to hydrolysis
    Macromolecules, 2009
    Co-Authors: Arlin L. Weikel, Lakshmi S. Nair, Nick R. Krogman, Cato T. Laurencin, Nhu Q Nguyen, Harry R. Allcock
    Abstract:

    The dipeptides alanyl-Glycine Ethyl Ester, valinyl-Glycine Ethyl Ester, and phenylalanyl-Glycine Ethyl Ester were synthesized by mixed anhydride solution-phase peptide reactions. The free N-terminus was used as a reactive site for nucleophilic replacement of the chlorine atoms in poly(dichlorophosphazene). The C-terminus was protected with an Ethyl Ester to prevent side reactions and cross-linking. The alanyl-Glycine Ethyl Ester replaced all the chlorine atoms in poly(dichlorophosphazene). However, replacement of all the chlorine atoms in poly(dichlorophosphazene) by valinyl-Glycine Ethyl Ester or phenylalanyl-Glycine Ethyl Ester polyphosphazenes was prevented by the insolubility of the partially substituted intermediates. To circumvent this problem, cosubstitution was carried out using the valinyl- or phenylalanyl Esters with Glycine Ethyl Ester or alanine Ethyl Ester in a 1:1 ratio. Cosubstituted polyphosphazenes with alanyl Glycine Ethyl Ester and Glycine Ethyl Ester or alanine Ethyl Ester were also sy...

  • synthesis of purine and pyrimidine containing polyphosphazenes physical properties and hydrolytic behavior
    Macromolecules, 2008
    Co-Authors: Nick R. Krogman, Lakshmi S. Nair, Cato T. Laurencin, Mark D Hindenlang, Harry R. Allcock
    Abstract:

    We report here the first examples of poly(organophosphazenes) with side groups derived from the purines guanine and adenine and the pyrimidine cytosine. Polymers with these purines or pyrimidines as the only side groups proved difficult to synthesize by macromolecular substitution techniques because of the insolubility of the intermediate products. Therefore, cosubstitution reactions of the poly(dichlorophosphazene) with Glycine Ethyl Ester, alanine Ethyl Ester, or diEthylene glycol mEthyl ether, followed by the respective purine or pyrimidine, were utilized. Each pair of side groups was incorporated into the polyphosphazene in a 1:1 ratio. 31P NMR spectroscopy verified the replacement of all the chlorine atoms, while 1H and 13C NMR techniques confirmed the presence and ratio of the different side group. DRIFT spectroscopy indicated that the attachment of the purines or pyrimidines was via the primary amino functionality. Glass transition temperatures ranged from −28 to −15 °C for the mixed-substituent po...

Michael L. Gross - One of the best experts on this subject based on the ideXlab platform.

  • isotope encoded carboxyl group footprinting for mass spectrometry based protein conformational studies
    Journal of the American Society for Mass Spectrometry, 2016
    Co-Authors: Hao Zhang, Michael L. Gross
    Abstract:

    We report an isotope-encoding method coupled with carboxyl-group footprinting to monitor protein conformational changes. The carboxyl groups of aspartic/glutamic acids and of the C-terminus of proteins can serve as reporters for protein conformational changes when labeled with Glycine Ethyl Ester (GEE) mediated by carbodiimide. In the new development, isotope-encoded “heavy” and “light” GEE are used to label separately the two states of the orange carotenoid protein (OCP) from cyanobacteria. Two samples are mixed (1:1 ratio) and analyzed by a single LC-MS/MS experiment. The differences in labeling extent between the two states are represented by the ratio of the “heavy” and “light” peptides, providing information about protein conformational changes. Combining isotope-encoded MS quantitative analysis and carboxyl-group footprinting reduces the time of MS analysis and improves the sensitivity of GEE and other footprinting.

  • mass spectrometry based protein footprinting characterizes the structures of oligomeric apolipoprotein e2 e3 and e4
    Biochemistry, 2011
    Co-Authors: Brian Gau, Kanchan Garai, Carl Frieden, Michael L. Gross
    Abstract:

    The three common isoforms of apolipoprotein E (ApoE) differ at two sites in their 299 amino acid sequence; these differences modulate the structure of ApoE to affect profoundly the isoform associations with disease. The e4 allele in particular is strongly associated with Alzheimer’s disease. The study of the structural effects of these mutation sites in aqueous media is hampered by the aggregation proclivity of each ApoE isoform. Hence, understanding the differences between isoforms has thus far relied on lower resolution biophysical measurements, mutagenesis, homology studies, and the use of truncated ApoE variants. In this study, we report two comparative studies of the ApoE family by using the mass spectrometry-based protein footprinting methods of FPOP and Glycine Ethyl Ester (GEE) labeling. The first experiment examines the three full-length WT isoforms in their tetrameric state and finds that the overall structures are similar, with the exception of M108 in ApoE4 which is more solvent-accessible in ...

  • carboxyl group footprinting maps the dimerization interface and phosphorylation induced conformational changes of a membrane associated tyrosine kinase
    Molecular & Cellular Proteomics, 2011
    Co-Authors: Hao Zhang, Don L Rempel, John Monsey, Ilan Vidavsky, Michael L. Gross, Wei Shen, Ron Bose
    Abstract:

    : Her4 is a transmembrane receptor tyrosine kinase belonging to the ErbB-EGFR family. It plays a vital role in the cardiovascular and nervous systems, and mutations in Her4 have been found in melanoma and lung cancer. The kinase domain of Her4 forms a dimer complex, called the asymmetric dimer, which results in kinase activation. Although a crystal structure of the Her4 asymmetric dimer is known, the dimer affinity and the effect of the subsequent phosphorylation steps on kinase domain conformation are unknown. We report here the use of carboxyl-group footprinting MS on a recombinant expressed, Her4 kinase-domain construct to address these questions. Carboxyl-group footprinting uses a water-soluble carbodiimide, 1-Ethyl-3-(3-dimEthylaminopropyl)carbodiimide, in the presence of Glycine Ethyl Ester, to modify accessible carboxyl groups on glutamate and aspartate residues. Comparisons of Her4 kinase-domain monomers versus dimers and of unphosphorylated versus phosphorylated dimers were made to map the dimerization interface and to determine phosphorylation induced-conformational changes. We detected 37 glutamate and aspartate residues that were modified, and we quantified their extents of modification by liquid chromatography MS. Five residues showed changes in carboxyl-group modification. Three of these residues are at the predicted dimer interface, as shown by the crystal structure, and the remaining two residues are on loops that likely have altered conformation in the kinase dimer. Incubating the Her4 kinase dimers with ATP resulted in dramatic increase in Tyr-850 phosphorylation, located on the activation loop, and this resulted in a conformational change in this loop, as evidenced by reduction in carboxyl-group modification. The kinase monomer-dimer equilibrium was measured using a titration format in which the extent of carboxyl-group footprinting was mathematically modeled to give the dimer association constant (1.5-6.8 × 10(12) dm(2)/mol). This suggests that the kinase-domain makes a significant contribution to the overall dimerization affinity of the full-length Her4 protein.

Peter T.m. Kenny - One of the best experts on this subject based on the ideXlab platform.

Lakshmi S. Nair - One of the best experts on this subject based on the ideXlab platform.

  • miscibility of choline substituted polyphosphazenes with plga and osteoblast activity on resulting blends
    Biomaterials, 2010
    Co-Authors: Arlin L. Weikel, Lakshmi S. Nair, Cato T. Laurencin, Steven G Owens, Nicole L Morozowich, Meng Deng, Harry R. Allcock
    Abstract:

    The preparation of phosphazene tissue engineering scaffolds with bioactive side groups has been accomplished using the biological buffer, choline chloride. Mixed-substituent phosphazene cyclic trimers (as model systems) and polymers with choline chloride and Glycine Ethyl Ester, alanine Ethyl Ester, valine Ethyl Ester, or phenylalanine Ethyl Ester were synthesized. Two different synthetic protocols were examined. A sodium hydride mediated route resulted in polyphosphazenes with a low choline content, while a cesium carbonate mediated process produced polyphosphazenes with higher choline content. The phosphazene structures and physical properties were studied using multinuclear NMR, differential scanning calorimetry (DSC), and gel permeation chromatography (GPC) techniques. The resultant polymers were then blended with PLGA (50:50) or PLGA (85:15) and characterized by DSC analysis and scanning electron microscopy (SEM). Polymer products obtained via the sodium hydride route produced miscible blends with both ratios of PLGA, while the cesium carbonate route yielded products with reduced blend miscibility. Heterophase hydrolysis experiments in aqueous media revealed that the polymer blends hydrolyzed to near-neutral pH media (∼5.8 to 6.8). The effect of different molecular structures on cellular adhesion showed osteoblast proliferation with an elevated osteoblast phenotype expression compared to PLGA over a 21-day culture period.

  • hydrogen bonding in blends of polyEsters with dipeptide containing polyphosphazenes
    Journal of Applied Polymer Science, 2010
    Co-Authors: Nick R. Krogman, Lakshmi S. Nair, Cato T. Laurencin, Arlin L. Weikel, Katherine A. Kristhart, Syam P. Nukavarapu, Nhu Q Nguyen, Harry R. Allcock
    Abstract:

    New biomedically erodible polymer composites were investigated. Polyphosphazenes containing the dipeptide side groups alanyl–Glycine Ethyl Ester, valinyl–Glycine Ethyl Ester, and phenylalanyl–Glycine Ethyl Ester were blended with poly(lactide-co-glycolide) (PLGA) with lactic to glycolic acid ratios of 50 : 50 [PLGA (50 : 50)] and 85 : 15 [PLGA (85 : 15)] with solution-phase techniques. Each dipeptide Ethyl Ester side group contains two NH protons that are capable of hydrogen bonding with the carbonyl functions of PLGA. Polyphosphazenes that contain only the dipeptide Ethyl Ester groups are insoluble in organic solvents and are thus unsuitable for solution-phase composite formation. To ensure solubility during and after synthesis, cosubstituted polymers with both dipeptide Ethyl Ester and Glycine or alanine Ethyl Ester side groups were used. Solution casting or electrospinning was used to fabricate polymer blend matrices with different ratios of polyphosphazene to polyEster, and their miscibilities were estimated with differential scanning calorimetry and scanning electron microscopy techniques. Polyphosphazenes with alanyl–Glycine Ethyl Ester side groups plus the second cosubstituent were completely miscible with PLGA (50 : 50) and PLGA (85 : 15) when processed via solution-casting techniques. This suggests that the hydrogen-bonding protons in alanyl–Glycine Ethyl Ester have access to the oxygen atoms of the carbonyl units in PLGA. However, when the same pair of polymers was electrospun from solution, the polymers proved to be immiscible. Solution-cast miscible polymer blends were obtained from PLGA (50 : 50) plus the polyphosphazene that was cosubstituted with valinyl–Glycine Ethyl Ester and Glycine Ethyl Ester side groups. © 2009 Wiley Periodicals, Inc. J Appl Polym Sci, 2010

  • polyphosphazenes that contain dipeptide side groups synthesis characterization and sensitivity to hydrolysis
    Macromolecules, 2009
    Co-Authors: Arlin L. Weikel, Lakshmi S. Nair, Nick R. Krogman, Cato T. Laurencin, Nhu Q Nguyen, Harry R. Allcock
    Abstract:

    The dipeptides alanyl-Glycine Ethyl Ester, valinyl-Glycine Ethyl Ester, and phenylalanyl-Glycine Ethyl Ester were synthesized by mixed anhydride solution-phase peptide reactions. The free N-terminus was used as a reactive site for nucleophilic replacement of the chlorine atoms in poly(dichlorophosphazene). The C-terminus was protected with an Ethyl Ester to prevent side reactions and cross-linking. The alanyl-Glycine Ethyl Ester replaced all the chlorine atoms in poly(dichlorophosphazene). However, replacement of all the chlorine atoms in poly(dichlorophosphazene) by valinyl-Glycine Ethyl Ester or phenylalanyl-Glycine Ethyl Ester polyphosphazenes was prevented by the insolubility of the partially substituted intermediates. To circumvent this problem, cosubstitution was carried out using the valinyl- or phenylalanyl Esters with Glycine Ethyl Ester or alanine Ethyl Ester in a 1:1 ratio. Cosubstituted polyphosphazenes with alanyl Glycine Ethyl Ester and Glycine Ethyl Ester or alanine Ethyl Ester were also sy...

  • synthesis of purine and pyrimidine containing polyphosphazenes physical properties and hydrolytic behavior
    Macromolecules, 2008
    Co-Authors: Nick R. Krogman, Lakshmi S. Nair, Cato T. Laurencin, Mark D Hindenlang, Harry R. Allcock
    Abstract:

    We report here the first examples of poly(organophosphazenes) with side groups derived from the purines guanine and adenine and the pyrimidine cytosine. Polymers with these purines or pyrimidines as the only side groups proved difficult to synthesize by macromolecular substitution techniques because of the insolubility of the intermediate products. Therefore, cosubstitution reactions of the poly(dichlorophosphazene) with Glycine Ethyl Ester, alanine Ethyl Ester, or diEthylene glycol mEthyl ether, followed by the respective purine or pyrimidine, were utilized. Each pair of side groups was incorporated into the polyphosphazene in a 1:1 ratio. 31P NMR spectroscopy verified the replacement of all the chlorine atoms, while 1H and 13C NMR techniques confirmed the presence and ratio of the different side group. DRIFT spectroscopy indicated that the attachment of the purines or pyrimidines was via the primary amino functionality. Glass transition temperatures ranged from −28 to −15 °C for the mixed-substituent po...

Cato T. Laurencin - One of the best experts on this subject based on the ideXlab platform.

  • miscibility of choline substituted polyphosphazenes with plga and osteoblast activity on resulting blends
    Biomaterials, 2010
    Co-Authors: Arlin L. Weikel, Lakshmi S. Nair, Cato T. Laurencin, Steven G Owens, Nicole L Morozowich, Meng Deng, Harry R. Allcock
    Abstract:

    The preparation of phosphazene tissue engineering scaffolds with bioactive side groups has been accomplished using the biological buffer, choline chloride. Mixed-substituent phosphazene cyclic trimers (as model systems) and polymers with choline chloride and Glycine Ethyl Ester, alanine Ethyl Ester, valine Ethyl Ester, or phenylalanine Ethyl Ester were synthesized. Two different synthetic protocols were examined. A sodium hydride mediated route resulted in polyphosphazenes with a low choline content, while a cesium carbonate mediated process produced polyphosphazenes with higher choline content. The phosphazene structures and physical properties were studied using multinuclear NMR, differential scanning calorimetry (DSC), and gel permeation chromatography (GPC) techniques. The resultant polymers were then blended with PLGA (50:50) or PLGA (85:15) and characterized by DSC analysis and scanning electron microscopy (SEM). Polymer products obtained via the sodium hydride route produced miscible blends with both ratios of PLGA, while the cesium carbonate route yielded products with reduced blend miscibility. Heterophase hydrolysis experiments in aqueous media revealed that the polymer blends hydrolyzed to near-neutral pH media (∼5.8 to 6.8). The effect of different molecular structures on cellular adhesion showed osteoblast proliferation with an elevated osteoblast phenotype expression compared to PLGA over a 21-day culture period.

  • hydrogen bonding in blends of polyEsters with dipeptide containing polyphosphazenes
    Journal of Applied Polymer Science, 2010
    Co-Authors: Nick R. Krogman, Lakshmi S. Nair, Cato T. Laurencin, Arlin L. Weikel, Katherine A. Kristhart, Syam P. Nukavarapu, Nhu Q Nguyen, Harry R. Allcock
    Abstract:

    New biomedically erodible polymer composites were investigated. Polyphosphazenes containing the dipeptide side groups alanyl–Glycine Ethyl Ester, valinyl–Glycine Ethyl Ester, and phenylalanyl–Glycine Ethyl Ester were blended with poly(lactide-co-glycolide) (PLGA) with lactic to glycolic acid ratios of 50 : 50 [PLGA (50 : 50)] and 85 : 15 [PLGA (85 : 15)] with solution-phase techniques. Each dipeptide Ethyl Ester side group contains two NH protons that are capable of hydrogen bonding with the carbonyl functions of PLGA. Polyphosphazenes that contain only the dipeptide Ethyl Ester groups are insoluble in organic solvents and are thus unsuitable for solution-phase composite formation. To ensure solubility during and after synthesis, cosubstituted polymers with both dipeptide Ethyl Ester and Glycine or alanine Ethyl Ester side groups were used. Solution casting or electrospinning was used to fabricate polymer blend matrices with different ratios of polyphosphazene to polyEster, and their miscibilities were estimated with differential scanning calorimetry and scanning electron microscopy techniques. Polyphosphazenes with alanyl–Glycine Ethyl Ester side groups plus the second cosubstituent were completely miscible with PLGA (50 : 50) and PLGA (85 : 15) when processed via solution-casting techniques. This suggests that the hydrogen-bonding protons in alanyl–Glycine Ethyl Ester have access to the oxygen atoms of the carbonyl units in PLGA. However, when the same pair of polymers was electrospun from solution, the polymers proved to be immiscible. Solution-cast miscible polymer blends were obtained from PLGA (50 : 50) plus the polyphosphazene that was cosubstituted with valinyl–Glycine Ethyl Ester and Glycine Ethyl Ester side groups. © 2009 Wiley Periodicals, Inc. J Appl Polym Sci, 2010

  • polyphosphazenes that contain dipeptide side groups synthesis characterization and sensitivity to hydrolysis
    Macromolecules, 2009
    Co-Authors: Arlin L. Weikel, Lakshmi S. Nair, Nick R. Krogman, Cato T. Laurencin, Nhu Q Nguyen, Harry R. Allcock
    Abstract:

    The dipeptides alanyl-Glycine Ethyl Ester, valinyl-Glycine Ethyl Ester, and phenylalanyl-Glycine Ethyl Ester were synthesized by mixed anhydride solution-phase peptide reactions. The free N-terminus was used as a reactive site for nucleophilic replacement of the chlorine atoms in poly(dichlorophosphazene). The C-terminus was protected with an Ethyl Ester to prevent side reactions and cross-linking. The alanyl-Glycine Ethyl Ester replaced all the chlorine atoms in poly(dichlorophosphazene). However, replacement of all the chlorine atoms in poly(dichlorophosphazene) by valinyl-Glycine Ethyl Ester or phenylalanyl-Glycine Ethyl Ester polyphosphazenes was prevented by the insolubility of the partially substituted intermediates. To circumvent this problem, cosubstitution was carried out using the valinyl- or phenylalanyl Esters with Glycine Ethyl Ester or alanine Ethyl Ester in a 1:1 ratio. Cosubstituted polyphosphazenes with alanyl Glycine Ethyl Ester and Glycine Ethyl Ester or alanine Ethyl Ester were also sy...

  • synthesis of purine and pyrimidine containing polyphosphazenes physical properties and hydrolytic behavior
    Macromolecules, 2008
    Co-Authors: Nick R. Krogman, Lakshmi S. Nair, Cato T. Laurencin, Mark D Hindenlang, Harry R. Allcock
    Abstract:

    We report here the first examples of poly(organophosphazenes) with side groups derived from the purines guanine and adenine and the pyrimidine cytosine. Polymers with these purines or pyrimidines as the only side groups proved difficult to synthesize by macromolecular substitution techniques because of the insolubility of the intermediate products. Therefore, cosubstitution reactions of the poly(dichlorophosphazene) with Glycine Ethyl Ester, alanine Ethyl Ester, or diEthylene glycol mEthyl ether, followed by the respective purine or pyrimidine, were utilized. Each pair of side groups was incorporated into the polyphosphazene in a 1:1 ratio. 31P NMR spectroscopy verified the replacement of all the chlorine atoms, while 1H and 13C NMR techniques confirmed the presence and ratio of the different side group. DRIFT spectroscopy indicated that the attachment of the purines or pyrimidines was via the primary amino functionality. Glass transition temperatures ranged from −28 to −15 °C for the mixed-substituent po...