Switchable Enzymatic Accessibility for Precision Cell‐Selective Surface Glycan Remodeling

Abstract

Precision cell-selective surface glycan remodeling is of vital importance for modulation of cell surface dynamics, tissue-specific imaging, and immunotherapy,b ut remains an unsolved challenge. Herein,w er eport as witchablee nzymatic accessibility (SEA)s trategy for highly specific editing of carbohydrate moieties of interesto nt he target cell surface. We demonstrate the blocking of enzyme in the inaccessible state with am etal-organic framework (MOF) cage and instantaneouss witching to the accessible state through disassembly of MOF.W efurthers how that this level of SEA regulation enablesi nitial guided enzyme deliveryt ot he target cell surface for subsequent cell-specific glycan remodeling, thus providingatemporally and spatially controlled tool for tuningt he glycosylation architectures. Terminal galactose/Nacetylgalactosamine( Gal/GalNAc) remodeling and terminal sialic acid (Sia) desialylation have been precisely achieved on targetc ells even with other cell lines in close spatial proximity.T he SEA protocolf eatures am odular and generically adaptabled esign, av ery short protocol duration (ca. 30 min or shorter), andavery high spatial resolving power( ability to differentiate immediately neighboring cell lines).

Aglycone sterics-selective enzymatic glycan remodeling

“…We treated MCF-7 cells with GO-PN (1∼15) at 4°C for 1 h, respectively. The hydroxyl group at the C6 position of terminal Gal/GalNAc on the cell surface can be oxidized to the aldehyde group ( Rannes et al., 2011 ), and then labeled with fluorescein-5-thiosemicarbazide (FTZ) to form a stable hydrazone linkage ( Zhang et al., 2019 ). The mean fluorescence intensity (MFI) of FTZ on GO-PN (1∼15) treated cells was normalized to MFI of equimolar GO-treated cells, to obtain a relative activity (%) for each type of GO-PNs.…”

Electrofluorochromic Imaging Analysis of Glycan Expression on Living Single Cell with Bipolar Electrode Arrays

“…At the same time, it also proved the successful sequential labeling process in the previous steps. Galactosyl groups had a certain amount of expression on the surface of normal liver cells, but the expression was significantly lower than that of liver cancer cells, which was in good agreement with the previous report …”

“…Galactosyl groups had a certain amount of expression on the surface of normal liver cells, but the expression was significantly lower than that of liver cancer cells, which was in good agreement with the previous report. 43 Evaluating Galactosyl Group evaluate the dynamic changes of galactosyl groups expression on the cell surface upon inhibitor treatment, 1 × 10 5 cells mL −1 HepG2 cells were treated with the inhibitors of N-glycan TM or O-glycan BG for 48 h, capturing a single cell in the anode trap, the recorded cathode EFC signal as shown in Figure 5C.…”

Glycoenzyme Tool Development: Principles, Screening Methods, and Recent Advances†

“…To ensure the spatial specificity, the enzyme is caged during the enzyme anchoring step, and then activated in situ to perform confined enzymatic oxidation. [ 129‐132 ] Utilization of these above‐mentioned methods will open up new possibilities in high‐throughput screening in glycoenzyme directed evolution.…”