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  1. Article ; Online: Differential Detection of O-GlcNAcylated proteins in the heart using antibodies

    Narayanan, Bhargavi / Zahra, Fiddia / Reeves, Russell A. / Aggarwal, Akanksha / O'Meally, Robert N. / Henry, Roger K. / Craven, Megan / Jacobson, Avital / Cole, Robert N. / Kohr, Mark J. / Umapathi, Priya / Zachara, Natasha E.

    Analytical Biochemistry. 2023 Oct., v. 678 p.115262-

    2023  

    Abstract: Thousands of mammalian intracellular proteins are dynamically modified by O-linked β−N-acetylglucosamine (O-GlcNAc). Global changes in O-GlcNAcylation have been associated with the development of cardiomyopathy, heart failure, hypertension, and ... ...

    Abstract Thousands of mammalian intracellular proteins are dynamically modified by O-linked β−N-acetylglucosamine (O-GlcNAc). Global changes in O-GlcNAcylation have been associated with the development of cardiomyopathy, heart failure, hypertension, and neurodegenerative disease. Levels of O-GlcNAc in cells and tissues can be detected using numerous approaches; however, immunoblotting using GlcNAc-specific antibodies and lectins is commonplace. The goal of this study was to optimize the detection of O-GlcNAc in heart lysates by immunoblotting. Using a combination of tissue fractionation, immunoblotting, and galactosyltransferase labeling, as well as hearts from wild-type and O-GlcNAc transferase transgenic mice, we demonstrate that contractile proteins in the heart are differentially detected by two commercially available antibodies (CTD110.6 and RL2). As CTD110.6 displays poor reactivity toward contractile proteins, and as these proteins represent a major fraction of the heart proteome, a better assessment of cardiac O-GlcNAcylation is obtained in total tissue lysates with RL2. The data presented highlight tissue lysis approaches that should aid the assessment of the cardiac O-GlcNAcylation by immunoblotting.
    Keywords cardiomyopathy ; fractionation ; genetically modified organisms ; heart ; heart failure ; hypertension ; immunoblotting ; lectins ; mammals ; neurodegenerative diseases ; proteome ; O-GlcNAc ; Glycosylation ; Antibodies ; Click chemistry
    Language English
    Dates of publication 2023-10
    Publishing place Elsevier Inc.
    Document type Article ; Online
    ZDB-ID 1110-1
    ISSN 1096-0309 ; 0003-2697
    ISSN (online) 1096-0309
    ISSN 0003-2697
    DOI 10.1016/j.ab.2023.115262
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  2. Article ; Online: Differential Detection of O-GlcNAcylated proteins in the heart using antibodies.

    Narayanan, Bhargavi / Zahra, Fiddia / Reeves, Russell A / Aggarwal, Akanksha / O'Meally, Robert N / Henry, Roger K / Craven, Megan / Jacobson, Avital / Cole, Robert N / Kohr, Mark J / Umapathi, Priya / Zachara, Natasha E

    Analytical biochemistry

    2023  Volume 678, Page(s) 115262

    Abstract: Thousands of mammalian intracellular proteins are dynamically modified by O-linked β-N-acetylglucosamine (O-GlcNAc). Global changes in O-GlcNAcylation have been associated with the development of cardiomyopathy, heart failure, hypertension, and ... ...

    Abstract Thousands of mammalian intracellular proteins are dynamically modified by O-linked β-N-acetylglucosamine (O-GlcNAc). Global changes in O-GlcNAcylation have been associated with the development of cardiomyopathy, heart failure, hypertension, and neurodegenerative disease. Levels of O-GlcNAc in cells and tissues can be detected using numerous approaches; however, immunoblotting using GlcNAc-specific antibodies and lectins is commonplace. The goal of this study was to optimize the detection of O-GlcNAc in heart lysates by immunoblotting. Using a combination of tissue fractionation, immunoblotting, and galactosyltransferase labeling, as well as hearts from wild-type and O-GlcNAc transferase transgenic mice, we demonstrate that contractile proteins in the heart are differentially detected by two commercially available antibodies (CTD110.6 and RL2). As CTD110.6 displays poor reactivity toward contractile proteins, and as these proteins represent a major fraction of the heart proteome, a better assessment of cardiac O-GlcNAcylation is obtained in total tissue lysates with RL2. The data presented highlight tissue lysis approaches that should aid the assessment of the cardiac O-GlcNAcylation by immunoblotting.
    MeSH term(s) Mice ; Animals ; Neurodegenerative Diseases ; Antibodies/metabolism ; Proteome/metabolism ; Heart ; Contractile Proteins/metabolism ; Acetylglucosamine ; Protein Processing, Post-Translational ; Mammals/metabolism
    Chemical Substances Antibodies ; Proteome ; Contractile Proteins ; Acetylglucosamine (V956696549)
    Language English
    Publishing date 2023-07-27
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
    ZDB-ID 1110-1
    ISSN 1096-0309 ; 0003-2697
    ISSN (online) 1096-0309
    ISSN 0003-2697
    DOI 10.1016/j.ab.2023.115262
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    In stock of ZB MED Cologne/Königswinter

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    Je nach Verfügbarkeit (siehe Angabe bei Bestand)
    bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular
    Jg. 1995 - 2021: Lesesall (1.OG)
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  3. Article: An investigation into FOXE1 polyalanine tract length in premature ovarian failure.

    Watkins, Wendy J / Harris, Sarah E / Craven, Megan J / Vincent, Andrea L / Winship, Ingrid M / Gersak, Ksenija / Shelling, Andrew N

    Molecular human reproduction

    2006  Volume 12, Issue 3, Page(s) 145–149

    Abstract: Premature ovarian failure (POF) is a common condition affecting 1% of women worldwide. There is strong evidence for genetic involvement in POF as many cases are familial, and mutations in several genes have been associated with POF. We investigated ... ...

    Abstract Premature ovarian failure (POF) is a common condition affecting 1% of women worldwide. There is strong evidence for genetic involvement in POF as many cases are familial, and mutations in several genes have been associated with POF. We investigated variation in FOXE1 polyalanine tract length, following the observation that polyalanine tract deletions are seen in the closely related FOXL2 in patients with POF. In addition, polyalanine tract expansions in FOXL2 are often seen in patients with blepharophimosis-ptosis-epicanthus inversus syndrome (BPES), a rare eyelid disorder often associated with POF. The FOXE1 polyalanine tract shows marked variation in its length between POF patients and normal controls, existing as an allele of 12, 14, 16, 17 or 19 alanine residues. We found evidence to suggest that variation in FOXE1 polyalanine tract length predisposes to POF.
    MeSH term(s) Adult ; Base Sequence ; Chromatography, High Pressure Liquid ; Female ; Forkhead Transcription Factors/genetics ; Genetic Predisposition to Disease/genetics ; Humans ; Molecular Sequence Data ; New Zealand ; Peptides/genetics ; Primary Ovarian Insufficiency/genetics ; Sequence Homology, Nucleic Acid ; Slovenia ; Trinucleotide Repeat Expansion/genetics
    Chemical Substances FOXE1 protein, human ; Forkhead Transcription Factors ; Peptides ; polyalanine (25191-17-7)
    Language English
    Publishing date 2006-03
    Publishing country England
    Document type Comparative Study ; Journal Article
    ZDB-ID 1324348-2
    ISSN 1460-2407 ; 1360-9947
    ISSN (online) 1460-2407
    ISSN 1360-9947
    DOI 10.1093/molehr/gal017
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    In stock of ZB MED Cologne/Königswinter

    Zs.A 4482: Show issues Location:
    Je nach Verfügbarkeit (siehe Angabe bei Bestand)
    bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular
    Jg. 1995 - 2021: Lesesall (2.OG)
    ab Jg. 2022: Lesesaal (EG)

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