Chemistry and mass spectrometry for protein structure determination in living cells
Abstract/Contents
- Abstract
- Protein structures speed discovery of life's molecular basis, but the artificial conditions of our structural techniques often disrupt natural protein behaviors. This could be sidestepped by chemically crosslinking adjacent residues together while a protein is still in its native cell. The protein could then be extracted and digested into mass spec--amenable peptides so the crosslinks can be seen by high-throughput mass spec. This would make a list of residue adjacencies useful to common structure determination software. Unfortunately, easily detectable crosslinkers are long and therefore vague about just how adjacent crosslinked residues are. This is because they facilitate detection by incorporating bulky moieties with distinctive behaviors during mass spec and purifiable tags. We show that a mass spec behavior ideal for crosslink detection can be encoded in a single well-placed atom in the crosslink. We also purify crosslinked peptides without adding anything to the crosslinker by inventing a method that isolates molecules with two peptidic N-termini. This frees us up to design crosslinking agents uncompromisingly tailored for high-resolution results. We synthesize these designs and investigate their utility.
Description
| Type of resource | text |
|---|---|
| Form | electronic resource; remote; computer; online resource |
| Extent | 1 online resource. |
| Place | California |
| Place | [Stanford, California] |
| Publisher | [Stanford University] |
| Copyright date | 2019; ©2019 |
| Publication date | 2019; 2019 |
| Issuance | monographic |
| Language | English |
Creators/Contributors
| Author | Wulff, Bjoern Erik | |
|---|---|---|
| Degree supervisor | Harbury, Pehr | |
| Thesis advisor | Harbury, Pehr | |
| Thesis advisor | Das, Rhiju | |
| Thesis advisor | Elias, Joshua | |
| Thesis advisor | Li, Lingyin | |
| Degree committee member | Das, Rhiju | |
| Degree committee member | Elias, Joshua | |
| Degree committee member | Li, Lingyin | |
| Associated with | Stanford University, Department of Biochemistry. |
Subjects
| Genre | Theses |
|---|---|
| Genre | Text |
Bibliographic information
| Statement of responsibility | Bjoern-Erik Wulff. |
|---|---|
| Note | Submitted to the Department of Biochemistry. |
| Thesis | Thesis Ph.D. Stanford University 2019. |
| Location | electronic resource |
Access conditions
- Copyright
- © 2019 by Bjoern Erik Wulff
- License
- This work is licensed under a Creative Commons Attribution Non Commercial 3.0 Unported license (CC BY-NC).
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