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Biocapture-Directed Chemical Labeling for Discerning Stressed States of Organelles.

Lei GaoYilong ShiEnkang ZhangJinxuan YouJiahuai HanXinhui SuShoufa Han
Published in: Analytical chemistry (2022)
Lysosomal rupture engaged in diverse diseases remains poorly discerned from lysosomal membrane permeabilization (LMP). We herein reported biocapture-directed chemical labeling (BCCL) for the discern of lysosomal rupture by tracking the release of optically labeled cathepsins from damaged lysosomes into the cytosol. BCCL entails covalent anchoring of an azide-tagged suicide substrate (Epo-LeuTyr Az ) to the enzyme active site and bioorthogonal ligation of the introduced azide with DBCO RC, a ratiometric sensor featuring an acidity-reporting red emissive X-rhodamine-lactam (ROX), blue emissive coumarin (CM) inert to pH, and DBCO reactive to azide. Aided with fluorescein isocyanate-labeled sialic acid (FITC-Sia), a probe remained in pH-elevated lysosomes but dissipated from LMP + lysosomes, BCCL enables optical discern of four states of lysosomes: ruptured lysosomes (blue in cytosol), LMP + lysosomes (blue in lysosomes), pH-elevated lysosomes (blue and green in lysosomes), and physiological lysosomes (blue, green and red in lysosomes). This approach could find applicability to study lysosome rupture over LMP in diseases and to evaluate lysosome rupture-inducing drugs.
Keyphrases
  • fluorescent probe
  • epstein barr virus
  • living cells
  • light emitting
  • subarachnoid hemorrhage
  • pet imaging
  • multidrug resistant
  • brain injury
  • pet ct
  • adverse drug