Protocols in Current Issue
    Radioactive Assay of in vitro Glutamylation Activity of the Legionella pneumophila Effector Protein SidJ
    Authors:  Alan G. Sulpizio, Jung-Ho Shin, Marena E. Minelli and Yuxin Mao, date: 10/05/2020, view: 294, Q&A: 0
    [Abstract] The Legionella effector protein SidJ has recently been identified to perform polyglutamylation on another Legionella effector, SdeA, ablating SdeA’s activity. SidJ is a kinase-like protein that requires the small eukaryotic protein calmodulin to perform glutamylation. Glutamylation is a relatively uncommon type of ...
    In vitro and in vivo Assessment of Protein Acetylation Status in Mycobacteria
    Authors:  Krishna K. Singh, Devendra P. Singh, Rambir Singh and Deepak K. Saini, date: 07/05/2019, view: 2201, Q&A: 0
    [Abstract] Protein acetylation is one of the standard post-translational modifications found in proteins across all organisms, along with phosphorylation which regulates diverse cellular processes. Acetylation of proteins can be enzymatically catalyzed through acetyltransferases, acetyl CoA synthetases or non-enzymatically through acyl carrier metabolic ...
    Characterization of Protein Domain Function via in vitro DNA Shuffling
    Authors:  Kathy Hiu Laam Po, Edward Wai Chi Chan and Sheng Chen, date: 06/05/2018, view: 3340, Q&A: 0
    [Abstract] We recently investigated the molecular events that drive evolution of the CTX-M-type β-lactamases by DNA shuffling of fragments of the blaCTX-M-14 and blaCTX-M-15 genes. Analysis of a total of 51 hybrid enzymes showed that enzymatic activity could be maintained in most cases, yet the enzymatically active ...
    Host-regulated Hepatitis B Virus Capsid Assembly in a Mammalian Cell-free System
    Authors:  Kuancheng Liu and Jianming Hu, date: 04/20/2018, view: 3766, Q&A: 0
    [Abstract] The hepatitis B virus (HBV) is an important global human pathogen and represents a major cause of hepatitis, liver cirrhosis and liver cancer. The HBV capsid is composed of multiple copies of a single viral protein, the capsid or core protein (HBc), plays multiple roles in the viral life cycle, and has emerged recently as a major target for ...
    ARP2/3 Phosphorylation Assay in the Presence of Recombinant Bacterial Effectors
    Authors:  Céline Michard, Lawrence L. LeClaire and Patricia Doublet, date: 04/05/2017, view: 5709, Q&A: 0
    [Abstract] The Actin-Related Protein 2/3 (ARP2/3) complex is an actin nucleator that generates a branched actin network in mammalian cells. In addition to binding nucleation promoting factors, LeClaire et al. demonstrated that its phosphorylation state is essential key for its activity (LeClaire et al., 2008). In cells, the ARP2/3 complex ...
    Extraction and Quantification of Polyphosphate in the Budding Yeast Saccharomyces cerevisiae
    Authors:  David Canadell, Samuel Bru, Josep Clotet and Joaquín Ariño, date: 07/20/2016, view: 8543, Q&A: 0
    [Abstract] Inorganic polyphosphate (polyP) is a linear polymer present in both prokaryotic and eukaryotic organisms and made from three to hundreds of orthophosphate residues linked by phosphoanhydride bonds. The biological role of this molecule goes beyond serving as Pi store or energy source to replace ATP. For instance, in yeast polyP levels have been ...
    In vitro Deneddylation Assay
    Authors:  Anna M. Köhler, Cindy Meister and Gerhard H. Braus, date: 03/20/2016, view: 6632, Q&A: 0
    [Abstract] Nedd8 is a small ubiquitin-like protein (9 kDa) covalently attached to a conserved lysine residue of a cullin protein which is part of cullin-RING ligases (CRLs). CRLs are major E3 ligases important for protein ubiquitination in the ubiquitin-proteasome pathway (UPP). The activity of CRLs is regulated by cycles of neddylation (CulA-N8, ~98 kDa) ...
    Detection of HBV C Protein Phosphorylation in the Cell
    Authors:  Jaesung Jung and Kyongmin Kim, date: 08/05/2015, view: 6016, Q&A: 0
    [Abstract] Among the seven serines and one threonine in the carboxyl-terminus of HBV C protein, all but one (serine 183) appear in the context of RxxS/T consensus phosphoacceptor motifs and also overlap with other consensus motifs, such as S/TP, RS, SPRRR, RRRS/T, or RRxS/T, suggesting that various cellular kinases phosphorylate these residues. To determine ...
    Biotinylation and Purification of Surface-exposed Helicobacter pylori Proteins
    Authors:  Bradley J. Voss and Timothy L. Cover, date: 04/20/2015, view: 6133, Q&A: 0
    [Abstract] Interactions between pathogenic bacteria and host cells are often mediated by proteins found on the surfaces of the bacteria. The Gram-negative bacterium Helicobacter pylori is predicted to produce at least 50 surface-exposed outer membrane proteins, but there has been relatively little progress in experimentally analyzing the ...
    Lectin Binding Analysis of Streptococcus mutans Glycoproteins
    Authors:  Alejandro Avilés-Reyes, José A. Lemos and Jacqueline Abranches, date: 04/05/2015, view: 6761, 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 ...

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