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Related Concept Videos

Protein Modifications in the RER01:26

Protein Modifications in the RER

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Modification of secretory and transmembrane proteins entering the rough ER begins in the ER lumen. These modifications aid in protein folding and stabilize the acquired tertiary structure. Protein modifications in the rough ER co-occur at different stages of protein folding.
Broadly, these modifications can be categorized into four main categories — glycosylation, formation of disulfide bonds, assembly of protein subunits, and specific proteolytic cleavages like removal of signal...
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Protein Glycosylation01:25

Protein Glycosylation

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Glycosylation, the most common post-translational modification for proteins, serves diverse functions. Adding sugars to proteins makes the proteins more resistant to proteolytic digestion. Glycosylated proteins can act as markers and receptors to promote cell-cell adhesion. Additionally, they have many essential quality control functions in the cell, such as correct protein folding and facilitating transport of misfolded proteins to the cytosol, which can be degraded.
Glycosylation occurs in...
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Oligosaccharide Assembly01:24

Oligosaccharide Assembly

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Protein glycosylation starts in the ER lumen and continues in the Golgi apparatus. Glycosyltransferases catalyze the addition of sugar molecules or glycosylation of proteins. Usually, these enzymes add sugars to the hydroxyl groups of selected serine or threonine residues to form O-linked glycans or the amino groups of asparagine residues to form N-linked glycans. Different positions on the same polypeptide chain can contain differently linked glycans.
Multiple sugar molecules that may or may...
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Protein Folding Quality Check in the RER01:29

Protein Folding Quality Check in the RER

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ER is the primary site for the maturation and folding of soluble and transmembrane secretory proteins. The calnexin cycle is a specific chaperone system that folds and assesses the confirmation of N-glycosylated proteins before they can exit the ER lumen. The primary players of this quality check pipeline are the lectins, ER-resident chaperones, and a glucosyl transferase enzyme. In case the calnexin system in the lumen fails to salvage a misfolded protein, it is transported to the cytoplasm...
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Coat Assembly and GTPases01:33

Coat Assembly and GTPases

4.0K
Vesicles incorporate different coat protein subunits in different cell locations, which changes the properties of the coat, such as the shape and geometry of the transport vesicles. Thus, vesicle coat proteins also play a significant role in cargo selection.
Coat assembly depends on the local availability of phosphatidylinositol phosphates or PIPs and GTP-binding proteins. Adaptor proteins, which link the coat proteins to the membrane, bind to these PIPs and play a crucial role in controlling...
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Tail-anchoring of Proteins in the ER Membrane01:45

Tail-anchoring of Proteins in the ER Membrane

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Tail-anchored, or TA, proteins are estimated to make up to 3-5% of membrane proteins found in the eukaryotic cell. Such proteins have a single transmembrane domain located approximately 30 amino acid residues upstream from the C-terminal end. As a result, the signal recognition particle (SRP) cannot guide a TA protein to the ER membrane for cotranslational insertion. Hence, they are integrated into the ER membrane post-translationally using their C-terminal end as the anchor. TA proteins...
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Detection of Protein S-Acylation using Acyl-Resin Assisted Capture
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Detection of Protein S-Acylation using Acyl-Resin Assisted Capture

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Stable Protein Sialylation in Physcomitrella.

Lennard L Bohlender1, Juliana Parsons1, Sebastian N W Hoernstein1

  • 1Plant Biotechnology, Faculty of Biology, University of Freiburg, Freiburg, Germany.

Frontiers in Plant Science
|January 4, 2021
PubMed
Summary
This summary is machine-generated.

This study successfully engineered moss (*Physcomitrium patens*) to perform human-like N-glycan sialylation on therapeutic proteins. This breakthrough enables plant-based production of complex glycoproteins with desired glycosylation patterns for medical use.

Keywords:
N-glycan humanizationN-glycan sialylationPMPglyco-engineeringglyco-optimizationplant-made pharmaceuticalsplant-made recombinant biopharmaceuticals

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Identification of Post-translational Modifications of Plant Protein Complexes
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Area of Science:

  • Biotechnology
  • Glycobiology
  • Molecular Biology

Background:

  • Recombinant glycoproteins are crucial for medical applications, with N-glycosylation profiles significantly impacting their function, structure, and safety.
  • The moss *Physcomitrium patens* (Physcomitrella) offers homogeneous complex-type N-glycosylation and meets biopharmaceutical standards, but lacks mammalian N-glycan sialylation.
  • Sialic acids are vital terminal modifications on human N-glycans, making their presence highly desirable for plant-based biopharmaceutical production.

Purpose of the Study:

  • To engineer *Physcomitrium patens* for the production of N-glycan sialylation on recombinant proteins.
  • To establish a plant-based system capable of synthesizing and attaching sialic acids to therapeutic glycoproteins.

Main Methods:

  • Generated a glyco-engineered moss line (Δxt/ft) lacking plant-specific sugar residues.
  • Introduced seven mammalian gene sequences for sialic acid synthesis, activation, transport, and transfer, with organelle-specific localization.
  • Modified endogenous galactosyltransferase activity by knocking out β1,3-GT and introducing a human β1,4-GT.

Main Results:

  • Successfully achieved stable N-glycan sialylation in the engineered moss line.
  • Confirmed production of free sialic acid (Neu5Ac) and its activated form (CMP-Neu5Ac).
  • Mass spectrometry verified functional complex-type N-glycan sialylation on a co-expressed recombinant human protein.

Conclusions:

  • Demonstrated the feasibility of N-glycan sialylation in *Physcomitrium patens* by integrating mammalian pathways.
  • This engineered moss line provides a novel platform for producing sialylated glycoproteins for therapeutic applications.
  • Overcomes a key limitation in plant-based glycoengineering, advancing the potential of plants as biopharmaceutical factories.