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Protein Glycosylation01:25

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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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Activity-Based Tracking of Glycan Turnover in Microbiomes.

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We developed a novel fluorescence resonance energy transfer (FRET) probe to visualize and quantify glycan metabolism in microbial communities. This tool tracks polysaccharide degradation, advancing microbiome research and understanding global carbon cycling.

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Area of Science:

  • Microbiology
  • Biochemistry
  • Environmental Science

Background:

  • Glycans are crucial components of microbial ecosystems, influencing the global carbon cycle and human health.
  • Current methods for tracking microbial glycan turnover and identifying active degraders are limited, especially without prior genomic information.

Purpose of the Study:

  • To develop and validate a novel activity-based fluorescence resonance energy transfer (FRET) probe for direct visualization and quantification of glycan metabolism in complex microbial communities.
  • To demonstrate the probe's utility in tracking α-mannan degradation, a key polysaccharide in algal blooms.

Main Methods:

  • Automated glycan assembly was used to synthesize a mannan hexasaccharide functionalized with a fluorescein-rhodamine FRET pair.
  • The FRET probe was validated using a recombinant endo-α-mannanase (GH76) in cell lysates, pure cultures, and complex microbiomes.
  • Microscopy and plate assays were employed to visualize and quantify in situ glycan turnover.

Main Results:

  • The FRET probe successfully enabled visualization and quantification of α-mannan degradation.
  • The probe demonstrated functionality across various biological contexts, from purified enzymes to complex marine microbiomes.
  • Spatiotemporal dynamics of α-mannan turnover were visualized in a marine microbial community.

Conclusions:

  • The developed glycan FRET probes are versatile tools for studying glycan metabolism.
  • These probes facilitate tracking of polysaccharide degradation across diverse biological scales, from enzymes to entire microbiomes.
  • This technology offers new possibilities for understanding microbial roles in biogeochemical cycles and host-microbe interactions.