Chemically tunable mucin chimeras assembled on living cells
Abstract
Mucins are a family of secreted and transmembrane glycoproteins characterized by a massive domain of dense O-glycosylation on serine and threonine residues. Mucins are intimately involved in immunity and cancer, yet elucidation of the biological roles of their glycodomains has been complicated by their massive size, domain polymorphisms, and variable glycosylation patterns. Here we developed a synthetic route to a library of compositionally defined, high-molecular weight, dual end-functionalized mucin glycodomain constructs via N-carboxyanhydride polymerization. These glycopolypeptides are the first synthetic analogs to our knowledge to feature the native α-GalNAc linkage to serine with molecular weights similar to native mucins, solving a nearly 50-year synthetic challenge. Physical characterization of the mimics revealed insights into the structure and properties of mucins. The synthetic glycodomains were end-functionalized with an optical probe and a tetrazine moiety, which allowed site-specific bioorthogonal conjugation to an engineered membrane protein on live mammalian cells. Lastly, this strategy in protein engineering will open avenues to explore the biological roles of cell surface mucins.
- Authors:
-
- Department of Chemistry, Stanford University, Stanford, CA 94305,
- Institute for Quantitative Biosciences, University of California, Berkeley, CA 94720,
- Institute for Quantitative Biosciences, University of California, Berkeley, CA 94720,, Department of Chemistry, University of California, Berkeley, CA 94720,, Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720,, Department of Physics, University of California, Berkeley, CA 94720,, Kavli Energy Nanosciences Institute at Berkeley, University of California, Berkeley, CA 94720,
- Department of Chemistry, Stanford University, Stanford, CA 94305,, Howard Hughes Medical Institute, Stanford University, Stanford, CA 94305
- Publication Date:
- Research Org.:
- University of California, Berkeley, CA (United States)
- Sponsoring Org.:
- USDOE
- OSTI Identifier:
- 1579282
- Alternate Identifier(s):
- OSTI ID: 1347690
- Grant/Contract Number:
- AC02-05CH11231; R01 GM059907
- Resource Type:
- Published Article
- Journal Name:
- Proceedings of the National Academy of Sciences of the United States of America
- Additional Journal Information:
- Journal Name: Proceedings of the National Academy of Sciences of the United States of America Journal Volume: 112 Journal Issue: 41; Journal ID: ISSN 0027-8424
- Publisher:
- Proceedings of the National Academy of Sciences
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; N-carboxyanhydride; glycoprotein; mucin; polypeptide; protein engineering
Citation Formats
Kramer, Jessica R., Onoa, Bibiana, Bustamante, Carlos, and Bertozzi, Carolyn R. Chemically tunable mucin chimeras assembled on living cells. United States: N. p., 2015.
Web. doi:10.1073/pnas.1516127112.
Kramer, Jessica R., Onoa, Bibiana, Bustamante, Carlos, & Bertozzi, Carolyn R. Chemically tunable mucin chimeras assembled on living cells. United States. https://doi.org/10.1073/pnas.1516127112
Kramer, Jessica R., Onoa, Bibiana, Bustamante, Carlos, and Bertozzi, Carolyn R. Tue .
"Chemically tunable mucin chimeras assembled on living cells". United States. https://doi.org/10.1073/pnas.1516127112.
@article{osti_1579282,
title = {Chemically tunable mucin chimeras assembled on living cells},
author = {Kramer, Jessica R. and Onoa, Bibiana and Bustamante, Carlos and Bertozzi, Carolyn R.},
abstractNote = {Mucins are a family of secreted and transmembrane glycoproteins characterized by a massive domain of dense O-glycosylation on serine and threonine residues. Mucins are intimately involved in immunity and cancer, yet elucidation of the biological roles of their glycodomains has been complicated by their massive size, domain polymorphisms, and variable glycosylation patterns. Here we developed a synthetic route to a library of compositionally defined, high-molecular weight, dual end-functionalized mucin glycodomain constructs via N-carboxyanhydride polymerization. These glycopolypeptides are the first synthetic analogs to our knowledge to feature the native α-GalNAc linkage to serine with molecular weights similar to native mucins, solving a nearly 50-year synthetic challenge. Physical characterization of the mimics revealed insights into the structure and properties of mucins. The synthetic glycodomains were end-functionalized with an optical probe and a tetrazine moiety, which allowed site-specific bioorthogonal conjugation to an engineered membrane protein on live mammalian cells. Lastly, this strategy in protein engineering will open avenues to explore the biological roles of cell surface mucins.},
doi = {10.1073/pnas.1516127112},
journal = {Proceedings of the National Academy of Sciences of the United States of America},
number = 41,
volume = 112,
place = {United States},
year = {Tue Sep 29 00:00:00 EDT 2015},
month = {Tue Sep 29 00:00:00 EDT 2015}
}
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https://doi.org/10.1073/pnas.1516127112
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