Final Report Summary - GLYCOTRACKER (Tracking Glycosylations with Targeted, Molecule-Sized “Noses”)
Glycobiology is poised to be the next revolution in biology and medicine; however, technical difficulties in detecting and characterizing glycans have prevented many biologists from entering this field, thus hampering new discoveries and innovations. This research program aimed at developing a conceptually novel technology that will allow straightforward identification of specific glycosylation patterns in biofluids and in live cells. We suggested that distinct glycosylation states will be differentiated by developing “artificial noses” the size of a single molecule, whereas selectivity toward specific glycoproteins can be obtained by attaching them to specific protein binders. To achieve high sensitivity and accuracy, several innovations in molecular recognition and fluorescence signaling have been integrated into the design of these unconventional molecular analytical devices.
Over the last years, we have validated the two main concepts underlying this proposal, namely, the ability to sense surfaces of specific proteins1-7 and the ability to discriminate among a vast number of different analytes by using unimolecular, pattern-generating fluorescent molecular probes.8-9 13 We have also shown how these two principles be integrated to afford a unimolecular fluorescent probe that can identify specific populations of protein biomarkers in biofluids and can discriminate among isoforms in living cells.10 The His-tag binders,1 protein surface receptors1, 4, and ‘turn-on’ fluorescent probes,3 which were suggested in our proposal, have been successfully developed. Based on these achievements, a functional glycosylation sensor that can analyze the glycosylation pattern of the Human chorionic gonadotropin (hCG) protein has already been developed and used to discriminate among glycoform mixtures.12 In addition to tracking glycosylation, the unique technologies that emerged from this research program have yielded new ideas and design principles that are relevant to various areas in chemistry. Developing pattern-generating molecular probes and devices,8-9 targeted, protein surface receptors,1-7 and biomimetics of signaling proteins4-6 are some of the original research directions that emerged from this study and that are being independently pursued.
1. Nissinkorn, Y.; Lahav-Mankovski, N.; Rabinkov, A.; Albeck, S.; Motiei, L.; Margulies, D., Sensing protein surfaces with targeted fluorescent receptors. Chem. Eur. J 2015, 21 (45), 15981-15987.
2. Nissinkorn, Y.; Motiei, L.; Margulies, D., Protein surface recognition with targeted fluorescent molecular probes. Receptors Clin. Investig. 2016, 3 : e1381. .
3. Unger-Angel, L.; Rout, B.; Ilani, T.; Eisenstein, M.; Motiei, L.; Margulies, D., Protein recognition by bivalent, 'turn-on' fluorescent molecular probes. Chem. Sci. 2015, 6 (10), 5419-5425.
4. Selvakumar, K.; Motiei, L.; Margulies, D., Enzyme−artificial enzyme interactions as a means for discriminating among structurally similar isozymes. J. Am. Chem. Soc 2015, 137 (15), 4892-4895.
5. Peri-Naor, R.; Ilani, T.; Motiei, L.; Margulies, D., Protein–protein communication and enzyme activation mediated by a synthetic chemical transducer. J. Am. Chem. Soc 2015, 137 (30), 9507-9510.
6. Peri-Naor, R.; Motiei, L.; Margulies, D., Artificial signal transduction therapy: a futuristic approach to disease treatment. Future Med. Chem. 2015, 7 (16), 2091-2093.
7. Peri-Naor, R.; Motiei, L.; Margulies, D., Mimicking the function of signaling proteins: toward artificial signal transduction therapy. J. Vis. Exp. 2016, 115 :e54396. .
8. Rout, B.; Motiei, L.; Margulies, D., Combinatorial fluorescent molecular sensors: the road to differential sensing at the molecular level. Synlett 2014, 25, 1050-1054.
9. Sarkar, T.; Selvakumar, K.; Motiei, L.; Margulies, D., Message in a moleule. Nat. Commun. 2016, 7, 11374-11382.
10. Pode, Z.; Peri-Naor, R.; Georgeson, J. M.; Ilani, T.; Kiss, V.; Unger, T.; Motiei, L.; Margulies, D., Protein recognition by a pattern-generating fluorescent molecular probe. Nat. Nanotechnol. 2017, 12, 1161-1168.
11. Hatai, J.; Motiei, L.; Margulies, D., Analyzing amyloid beta aggregates with a combinatorial fluorescent molecular sensor. J. Am. Chem. Soc. 2017, 139 (6), 2136-2139.
12. Peri-Naor, R.; Motiei, L.; Margulies, D., Tracking glycosylation by a self-assembled, pattern generating fluorescent molecular probe. In preparation 2018.
13. Lustgarten, O.; Carmieli, R.; Motiei, L.; Margulies, D., A Molecular Secret Sharing Scheme. Angew. Chem. Int. Ed. in press. 2018.
Over the last years, we have validated the two main concepts underlying this proposal, namely, the ability to sense surfaces of specific proteins1-7 and the ability to discriminate among a vast number of different analytes by using unimolecular, pattern-generating fluorescent molecular probes.8-9 13 We have also shown how these two principles be integrated to afford a unimolecular fluorescent probe that can identify specific populations of protein biomarkers in biofluids and can discriminate among isoforms in living cells.10 The His-tag binders,1 protein surface receptors1, 4, and ‘turn-on’ fluorescent probes,3 which were suggested in our proposal, have been successfully developed. Based on these achievements, a functional glycosylation sensor that can analyze the glycosylation pattern of the Human chorionic gonadotropin (hCG) protein has already been developed and used to discriminate among glycoform mixtures.12 In addition to tracking glycosylation, the unique technologies that emerged from this research program have yielded new ideas and design principles that are relevant to various areas in chemistry. Developing pattern-generating molecular probes and devices,8-9 targeted, protein surface receptors,1-7 and biomimetics of signaling proteins4-6 are some of the original research directions that emerged from this study and that are being independently pursued.
1. Nissinkorn, Y.; Lahav-Mankovski, N.; Rabinkov, A.; Albeck, S.; Motiei, L.; Margulies, D., Sensing protein surfaces with targeted fluorescent receptors. Chem. Eur. J 2015, 21 (45), 15981-15987.
2. Nissinkorn, Y.; Motiei, L.; Margulies, D., Protein surface recognition with targeted fluorescent molecular probes. Receptors Clin. Investig. 2016, 3 : e1381. .
3. Unger-Angel, L.; Rout, B.; Ilani, T.; Eisenstein, M.; Motiei, L.; Margulies, D., Protein recognition by bivalent, 'turn-on' fluorescent molecular probes. Chem. Sci. 2015, 6 (10), 5419-5425.
4. Selvakumar, K.; Motiei, L.; Margulies, D., Enzyme−artificial enzyme interactions as a means for discriminating among structurally similar isozymes. J. Am. Chem. Soc 2015, 137 (15), 4892-4895.
5. Peri-Naor, R.; Ilani, T.; Motiei, L.; Margulies, D., Protein–protein communication and enzyme activation mediated by a synthetic chemical transducer. J. Am. Chem. Soc 2015, 137 (30), 9507-9510.
6. Peri-Naor, R.; Motiei, L.; Margulies, D., Artificial signal transduction therapy: a futuristic approach to disease treatment. Future Med. Chem. 2015, 7 (16), 2091-2093.
7. Peri-Naor, R.; Motiei, L.; Margulies, D., Mimicking the function of signaling proteins: toward artificial signal transduction therapy. J. Vis. Exp. 2016, 115 :e54396. .
8. Rout, B.; Motiei, L.; Margulies, D., Combinatorial fluorescent molecular sensors: the road to differential sensing at the molecular level. Synlett 2014, 25, 1050-1054.
9. Sarkar, T.; Selvakumar, K.; Motiei, L.; Margulies, D., Message in a moleule. Nat. Commun. 2016, 7, 11374-11382.
10. Pode, Z.; Peri-Naor, R.; Georgeson, J. M.; Ilani, T.; Kiss, V.; Unger, T.; Motiei, L.; Margulies, D., Protein recognition by a pattern-generating fluorescent molecular probe. Nat. Nanotechnol. 2017, 12, 1161-1168.
11. Hatai, J.; Motiei, L.; Margulies, D., Analyzing amyloid beta aggregates with a combinatorial fluorescent molecular sensor. J. Am. Chem. Soc. 2017, 139 (6), 2136-2139.
12. Peri-Naor, R.; Motiei, L.; Margulies, D., Tracking glycosylation by a self-assembled, pattern generating fluorescent molecular probe. In preparation 2018.
13. Lustgarten, O.; Carmieli, R.; Motiei, L.; Margulies, D., A Molecular Secret Sharing Scheme. Angew. Chem. Int. Ed. in press. 2018.