Fluorometric visualization of mucin 1 glycans on cell surfaces based on rolling-mediated cascade amplification and CdTe quantum dots
- 112 Downloads
A rolling-mediated cascade (RMC) amplification strategy is described for improved visualization of profiling glycans of mucin 1 (MUC 1) on cell surfaces. CdTe quantum dots (QDs) are used as fluorescent labels. The RMC based amplification allows even distinct glycoforms of MUC1 to be visualized on the surface of MCF-7 cell via an amplified Förster resonance energy transfer (FRET) imaging strategy that works at excitation/emission wavelengths of 345/610 nm. This is achieved by utilizing antibody against MUC1 modified with the fluorescent label 7-amino-4-methylcoumarin-3-acetic acid (AMCA) as the energy donor in FRET. The QDs (used to label surface glycans) act as acceptors. N-Azidoacetylgalactosamine-Acetylated (Ac4GalNAz) as a non-natural azido sugar, can be incorporated into the glycans of the cell surface, which can promote further labeling. The method has the advantage of only requiring a small amount of non-natural sugar to be introduced in metabolic glycan labeling since too much of an artificial sugar will interfere with the physiological functions of cells.
KeywordsGlycosylation Azide polysaccharide Metabolic labeling Glycoprotein Cancer marker Quantum dots DNA probe Click reaction Rolling-mediated cascade amplification FRET
We sincerely appreciate the National Natural Science Foundation of China for the financial support (81573388). This work was supported by “Qing Lan Project of Jiangsu province” and “Six talent peaks project of Jiangsu Province (YY-032)”. This work was also supported by the Open Project Program of Jiangsu Key Laboratory for Pharmacology and Safety Evaluation of Chinese Materia Medica (No. JKLPSE201805) and the Project of the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD).
Compliance with ethical standards
Conflict of interest
The author(s) declare that they have no competing interests.
- 1.Stowell SR, Ju T, Cummings RD (2015) Protein glycosylation in cancer. Annu Rev Pathol 10(1):473–510. https://doi.org/10.1146/annurev-pathol-012414-040438 CrossRefPubMedPubMedCentralGoogle Scholar
- 3.Paszek MJ, Dufort CC, Rossier O, Bainer R, Mouw JK, Godula K, Hudak JE, Lakins JN, Wijekoon AC, Cassereau L, Rubashkin MG, Magbanua MJ, Thorn KS, Davidson MW, Rugo HS, Park JW, Hammer DA, Giannone G, Bertozzi CR, Weaver VM (2014) The cancer glycocalyx mechanically primes integrin-mediated growth and survival. Nature 511(7509):319–325. https://doi.org/10.1038/nature13535 CrossRefPubMedPubMedCentralGoogle Scholar
- 6.Katorcha E, Makarava N, Savtchenko R, Baskakov IV (2014) Sialylation of prion protein controls the rate of prion amplification, the cross-species barrier, the ratio of PrPSc glycoform and prion infectivity. PLoS Pathog 10:e1004366. https://doi.org/10.1371/journal.ppat.1004366 CrossRefPubMedPubMedCentralGoogle Scholar
- 24.Grünberg R, Burnier JV, Ferrar T, Beltran-Sastre V, Stricher F, Sloot AMVD, Garcia-Olivas R, Mallabiabarrena A, Sanjuan X, Zimmermann T, Serrano L (2013) Engineering of weak helper interactions for high-efficiency FRET probes. Nature Method 10(10):1021–1027. https://doi.org/10.1038/NMETH.2625 CrossRefGoogle Scholar
- 29.Zhu D, Miao ZY, Hu Y, Zhang XJ (2018) Single-step, homogeneous and sensitive detection for microRNAs with dual recognition steps based on luminescence resonance energy transfer (LRET) using upconversion nanoparticles. Biosens Bioelectron 100:475–481. https://doi.org/10.1016/j.bios.2017.09.039 CrossRefPubMedGoogle Scholar
- 31.Rong J, Han J, Dong L, Tan Y, Yang H, Feng L, Wang Q, Meng R, Zhao J, Wang S, Chen X (2014) Glycan imaging in intact rat hearts and glycoproteomic analysis reveal the upregulation of sialylation during cardiac hypertrophy. J Am Chem Soc 136(50):17468–17476. https://doi.org/10.1021/ja508484c CrossRefPubMedGoogle Scholar