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Seiji Shinkai - One of the best experts on this subject based on the ideXlab platform.

  • polysaccharide polynucleotide complexes 2 complementary polynucleotide mimic behavior of the natural polysaccharide schizophyllan in the macromolecular complex with single stranded rna and dna
    Biomacromolecules, 2001
    Co-Authors: Kazuo Sakurai, Masami Mizu, Seiji Shinkai
    Abstract:

    Schizophyllan is an extracellular polysaccharide consisting of a β-1,3-d-glucan main chain and exists as a triple helix in water and as a single chain in dimethyl sulfoxide (DMSO). When the single chain of schizophyllan (s-SPG) was mixed with poly(C), poly(A), poly(dA), or poly(dT), they form a macromolecular complex. On the other hand, poly(G), poly(U), poly(I), poly(dG), and poly(dC) do not. This nucleotide specificity evidences that the hydrogen bonds are essential to form the complex, because the former nucleotides have an unoccupied hydrogen-Bonding Site and the latter ones use the hydrogen-Bonding Sites in the intramolecular aggregation (i.e., such as the G quartet for poly(G) and poly(dG) and the U hairpin for poly(U)). The hypochromic effect and the increment in the circular dichroism (CD) intensity are observed in accordance with the complex formation. These facts indicate that the base stacking is enhanced in the complex. The solvent−composition (DMSO/water) dependence demonstrates that the hydr...

  • polysaccharide polynucleotide complexes 2 complementary polynucleotide mimic behavior of the natural polysaccharide schizophyllan in the macromolecular complex with single stranded rna and dna
    Biomacromolecules, 2001
    Co-Authors: Kazuo Sakurai, Masami Mizu, Seiji Shinkai
    Abstract:

    Schizophyllan is an extracellular polysaccharide consisting of a beta-1,3-D-glucan main chain and exists as a triple helix in water and as a single chain in dimethyl sulfoxide (DMSO). When the single chain of schizophyllan (s-SPG) was mixed with poly(C), poly(A), poly(dA), or poly(dT), they form a macromolecular complex. On the other hand, poly(G), poly(U), poly(I), poly(dG), and poly(dC) do not. This nucleotide specificity evidences that the hydrogen bonds are essential to form the complex, because the former nucleotides have an unoccupied hydrogen-Bonding Site and the latter ones use the hydrogen-Bonding Sites in the intramolecular aggregation (i.e., such as the G quartet for poly(G) and poly(dG) and the U hairpin for poly(U)). The hypochromic effect and the increment in the circular dichroism (CD) intensity are observed in accordance with the complex formation. These facts indicate that the base stacking is enhanced in the complex. The solvent-composition (DMSO/water) dependence demonstrates that the hydrophobic interaction is important to form the complex as well as the hydrogen-Bonding interaction. With increasing temperature the complex dissociates cooperatively and the melting curve enables the thermodynamic parameters to be evaluated (delta H = -60 to 70 kcal mol-1 and delta S = -150 to 200 cal mol-1 K-1). These values are comparable with those for double helix DNA. Namely, the complex can be characterized by enhancement of the base stacking, cooperative dissociation, the similar thermodynamic parameters to DNA, and combination of the hydrogen-Bonding and hydrophobic interactions to form the higher-order structure. These facts surprisingly coincide with characters of the double helix of DNA. In other words, the s-SPG molecule behaves as if it were a complementary polynucleotide chain for the corresponding polynucleotide. Furthermore, stoichiometric study suggested that the complex structure is a triple helix consisting of two s-SPG and one poly(C) or poly(A) chains.

Kazuo Sakurai - One of the best experts on this subject based on the ideXlab platform.

  • polysaccharide polynucleotide complexes 2 complementary polynucleotide mimic behavior of the natural polysaccharide schizophyllan in the macromolecular complex with single stranded rna and dna
    Biomacromolecules, 2001
    Co-Authors: Kazuo Sakurai, Masami Mizu, Seiji Shinkai
    Abstract:

    Schizophyllan is an extracellular polysaccharide consisting of a β-1,3-d-glucan main chain and exists as a triple helix in water and as a single chain in dimethyl sulfoxide (DMSO). When the single chain of schizophyllan (s-SPG) was mixed with poly(C), poly(A), poly(dA), or poly(dT), they form a macromolecular complex. On the other hand, poly(G), poly(U), poly(I), poly(dG), and poly(dC) do not. This nucleotide specificity evidences that the hydrogen bonds are essential to form the complex, because the former nucleotides have an unoccupied hydrogen-Bonding Site and the latter ones use the hydrogen-Bonding Sites in the intramolecular aggregation (i.e., such as the G quartet for poly(G) and poly(dG) and the U hairpin for poly(U)). The hypochromic effect and the increment in the circular dichroism (CD) intensity are observed in accordance with the complex formation. These facts indicate that the base stacking is enhanced in the complex. The solvent−composition (DMSO/water) dependence demonstrates that the hydr...

  • polysaccharide polynucleotide complexes 2 complementary polynucleotide mimic behavior of the natural polysaccharide schizophyllan in the macromolecular complex with single stranded rna and dna
    Biomacromolecules, 2001
    Co-Authors: Kazuo Sakurai, Masami Mizu, Seiji Shinkai
    Abstract:

    Schizophyllan is an extracellular polysaccharide consisting of a beta-1,3-D-glucan main chain and exists as a triple helix in water and as a single chain in dimethyl sulfoxide (DMSO). When the single chain of schizophyllan (s-SPG) was mixed with poly(C), poly(A), poly(dA), or poly(dT), they form a macromolecular complex. On the other hand, poly(G), poly(U), poly(I), poly(dG), and poly(dC) do not. This nucleotide specificity evidences that the hydrogen bonds are essential to form the complex, because the former nucleotides have an unoccupied hydrogen-Bonding Site and the latter ones use the hydrogen-Bonding Sites in the intramolecular aggregation (i.e., such as the G quartet for poly(G) and poly(dG) and the U hairpin for poly(U)). The hypochromic effect and the increment in the circular dichroism (CD) intensity are observed in accordance with the complex formation. These facts indicate that the base stacking is enhanced in the complex. The solvent-composition (DMSO/water) dependence demonstrates that the hydrophobic interaction is important to form the complex as well as the hydrogen-Bonding interaction. With increasing temperature the complex dissociates cooperatively and the melting curve enables the thermodynamic parameters to be evaluated (delta H = -60 to 70 kcal mol-1 and delta S = -150 to 200 cal mol-1 K-1). These values are comparable with those for double helix DNA. Namely, the complex can be characterized by enhancement of the base stacking, cooperative dissociation, the similar thermodynamic parameters to DNA, and combination of the hydrogen-Bonding and hydrophobic interactions to form the higher-order structure. These facts surprisingly coincide with characters of the double helix of DNA. In other words, the s-SPG molecule behaves as if it were a complementary polynucleotide chain for the corresponding polynucleotide. Furthermore, stoichiometric study suggested that the complex structure is a triple helix consisting of two s-SPG and one poly(C) or poly(A) chains.

Thorsten Bach - One of the best experts on this subject based on the ideXlab platform.

Tsong-song Hwang - One of the best experts on this subject based on the ideXlab platform.

Eric S Simon - One of the best experts on this subject based on the ideXlab platform.

  • the use of hydrolytic enzymes and multi stage tandem mass spectrometry to analyze pyridoxal phosphate modified peptides
    Analytical Biochemistry, 2019
    Co-Authors: Eric S Simon, Phil Andrews
    Abstract:

    Abstract A previous approach was established that allowed direct identification of pyridoxal-5ˊ-phosphate (PLP) Bonding Sites in proteins using mass spectrometry after tryptic proteolysis. The approach required peptide mass fingerprinting owing to suppressed amide backbone fragmentation in favor of side-chain elimination of diagnostic product ions from PLP-derivatized lysyl residues. While sufficient for purified proteins, unambiguous sequence determination is needed to assign PLP Bonding Sites in unknown proteins in complex mixtures. Here, we describe the use of hydrolytic enzymes and multi-stage tandem mass spectrometry to elucidate the amino acid sequence and PLP Bonding Site in PLP-modified peptides.

  • determination of pyridoxal 5 phosphate plp Bonding Sites in proteins a peptide mass fingerprinting approach based on diagnostic tandem mass spectral features of plp modified peptides
    Rapid Communications in Mass Spectrometry, 2009
    Co-Authors: Eric S Simon, John Allison
    Abstract:

    Peptides modified by pyridoxal-5'-phosphate (PLP), linked to a lysine residue via reductive amination, exhibit distinct spectral characteristics in the collision-induced dissociation (CID) tandem mass (MS/MS) spectra that are described here. The MS/MS spectra typically display two dominant peaks whose m/z values correspond to neutral losses of [H3PO4] (-98 Da) and the PLP moiety as [C8H10NO5P] (-231 Da) from the precursor peptide ion, respectively. Few other peaks are observed. Recognition of this distinct fragmentation behavior is imperative since determining sequences and Sites of modifications relies on the formation of amide backbone cleavage products for subsequent interpretation via proteome database searching. Additionally, PLP-modified peptides exhibit suppressed precursor ionization efficiency which diminishes their detection in complex mixtures. Presented here is a protocol which describes an enrichment strategy for PLP-modified peptides combined with neutral loss screening and peptide mass fingerprinting to map the PLP-Bonding Site in a known PLP-dependent protein. This approach represents an efficient alternative to Site-directed mutagenesis which has been the traditional method used for PLP-Bonding Site localization in proteins.