Protocols in Current Issue
    PNGase Sensitivity Assay to Study the Folding Status of Proteins
    Authors:  Satoshi Ninagawa and Kazutoshi Mori, date: 10/05/2016, view: 2514, Q&A: 0
    [Abstract] This protocol aims to evaluate folding status of proteins, utilizing peptide:N-glycanase (PNGase) sensitivity. In the cytosol, PNGase works as a deglycosylation-enzyme. N-glycans on unfolded/misfolded proteins are more susceptible to PNGase than N-glycans on folded proteins because of the preference of PNGase to non-native proteins. PNGase is ...
    LC/MS-based Detection of Hydroxyproline O-galactosyltransferase Activity
    Authors:  Mari Ogawa-Ohnishi and Yoshikatsu Matsubayashi, date: 01/20/2016, view: 2885, Q&A: 0
    [Abstract] Arabinogalactan proteins (AGPs) are plant-specific extracellular glycoproteins regulating a variety of processes during growth and development. AGP biosynthesis involves O-galactosylation of hydroxyproline (Hyp) residues followed by a stepwise elongation of the complex sugar chains. The initial Hyp O-galactosylation is mediated ...
    Preparation and Analysis of Crude Autolytic Enzyme Extracts from Staphylococcus aureus
    Authors:  Filipa Vaz and Sérgio R. Filipe, date: 12/20/2015, view: 5201, Q&A: 0
    [Abstract] The metabolism of the cell surface during bacterial cell division involves synthesis and degradation of peptidoglycan (PGN), the major component of the bacterial cell wall. Bacteria have to ensure that their surface remains capable of withstanding high turgor pressures and, simultaneously, that the PGN at their surface is concealed from receptors ...
    Lectin Binding Analysis of Streptococcus mutans Glycoproteins
    Authors:  Alejandro Avilés-Reyes, José A. Lemos and Jacqueline Abranches, date: 04/05/2015, view: 3772, Q&A: 0
    [Abstract] Bacterial glycoproteins are of increasing interest due to their abundance in nature and importance in health and infectious diseases. However, only a very small fraction of bacterial glycoproteins have been characterized and its post-translational modification machinery identified. While analysis of glycoproteins can be achieved through various ...
    Dimethylmethylene Blue Assay (DMMB)
    Authors:  Vivien Jane Coulson- Thomas and Tarsis Ferreira Gesteira, date: 09/20/2014, view: 26821, Q&A: 6
    [Abstract] Glycosaminoglycans (GAGs) are long unbranched polysaccharides consisting of repeating disaccharide units composed of a hexosamine (glucosamine or galactosamine) and a hexuronic acid (glucuronic or iduronic acid). Depending on the disaccharide unit the GAGs can be organized into five groups: chondroitin sulfate, dermatan sulfate, heparan sulfate, ...
    Determination of D-galactofuranose Content of Galactomannoproteins in Aspergillus nidulans
    [Abstract] Galactofuranose (Galf) is a component of several polysaccharides and glycoconjugates in certain species of filamentous fungi. Galf residues are frequently found in Aspergillus glycoproteins, including N-glycans and O-mannose glycans that modify many cell wall proteins and extracellular enzymes. It is ...
    Seed Coat Ruthenium Red Staining Assay
    Authors:  Heather E. McFarlane, Delphine Gendre and Tamara L. Western, date: 04/05/2014, view: 8937, Q&A: 0
    [Abstract] The goal of this protocol is to assay for defects in synthesis/secretion/release of seed coat mucilage by ruthenium red staining of mature whole seeds.

    The mucilage secretory cells of the Arabidopsis seed coat synthesize and secrete a large quantity of primarily pectinaceous mucilage to a ring-shaped apical domain during their ...
    Preparation of Arabinogalactan Glycoproteins from Plant Tissue
    Author:  Derek T. A. Lamport, date: 10/05/2013, view: 5445, Q&A: 1
    [Abstract] This supplements an earlier protocol (Popper, 2011) for the extraction and assay of cell surface arabinogalactan proteins (AGPs). These highly glycosylated glycoproteins (~95% carbohydrate) contain numerous glycomodules with paired glucuronic acid residues that bind Ca2+ in a pH dependent manner (Lamport and Varnai, 2013). Classical ...