Journal
/
Immunology and Infection
/
Targeting Cysteine Thiols for in Vitro Site-specific Glycosylation of Recombinant Proteins
JoVE Journal
Immunology and Infection
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
JoVE Journal Immunology and Infection
Targeting Cysteine Thiols for in Vitro Site-specific Glycosylation of Recombinant Proteins

Targeting Cysteine Thiols for in Vitro Site-specific Glycosylation of Recombinant Proteins

DOI:
10.3791/56302-v

•

11:25 min

•

October 04, 2017

•

, , , , ,
1Department of Physiology and Pharmacology, Schulich School of Medicine and Dentistry,University of Western Ontario

Chapters

  • 00:05Title
  • 01:09Polymerase Chain Reaction (PCR)-mediated Site-directed Mutagenesis
  • 03:44Uniform 15N-labeled Protein Expression in BL21 ΔE3 E. coli
  • 05:24Purification of Recombinant Protein from E. coli
  • 07:50Chemical Attachment of Glucose-5-MTS to Protein by Dialysis
  • 08:48Results: Site-selective Cys Mutation Followed by In Vitro Glycosylation in STIM1 EF-SAM Protein
  • 10:11Conclusion

Summary

Automatic Translation

Automatic Translation

Biochemical and structural analyses of glycosylated proteins require relatively large amounts of homogeneous samples. Here, we present an efficient chemical method for site-specific glycosylation of recombinant proteins purified from bacteria by targeting reactive Cys thiols.

Tags

Cysteine ThiolsSite-specific GlycosylationRecombinant ProteinsStructural BiologyGlycosylation PatternsDisease TherapyPCR MutagenesisDPN1 DigestionProtein Expression

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

Generation of Recombinant Arenavirus for Vaccine Development in FDA-Approved Vero Cells

Expression of Functional Recombinant Hemagglutinin and Neuraminidase Proteins from the Novel H7N9 Influenza Virus Using the Baculovirus Expression System

Monitoring the Assembly of a Secreted Bacterial Virulence Factor Using Site-specific Crosslinking

Intranasal Administration of Recombinant Influenza Vaccines in Chimeric Mouse Models to Study Mucosal Immunity

Evaluation of the Efficacy And Toxicity of RNAs Targeting HIV-1 Production for Use in Gene or Drug Therapy

Murine Lymphocyte Labeling by 64Cu-Antibody Receptor Targeting for In Vivo Cell Trafficking by PET/CT

System for Efficacy and Cytotoxicity Screening of Inhibitors Targeting Intracellular Mycobacterium tuberculosis

In Vitro Methods for Comparing Target Binding and CDC Induction Between Therapeutic Antibodies: Applications in Biosimilarity Analysis

Generating Recombinant Avian Herpesvirus Vectors with CRISPR/Cas9 Gene Editing

Separation of Rat Epidermis and Dermis with Thermolysin to Detect Site-Specific Inflammatory mRNA and Protein

Read Article