Enabling Structural Proteomics with High Efficiency Protein Enrichment Technology

Raval, Shaunak

Enabling Structural Proteomics with High Efficiency Protein Enrichment Technology

dc.contributor.advisor	Schriemer, David C.
dc.contributor.author	Raval, Shaunak
dc.contributor.committeemember	MacCallum, Justin L.
dc.contributor.committeemember	Osthoff, Hans D.
dc.date	2023-11
dc.date.accessioned	2023-09-26T14:15:13Z
dc.date.available	2023-09-26T14:15:13Z
dc.date.issued	2023-09-13
dc.description.abstract	The functional state of proteins is inherently flexible, which allows them to interact with other biomolecules, including other proteins, to carry out many of their cellular functions. Understanding the structural dynamics of proteins and their network of associations is key to understanding their role in biology. Proteomics, the collection of mass spectrometry (MS)-based techniques to study proteins, provides a broad view of the organization of protein structure, from an individual dynamic unit to large-scale multiprotein assemblies, enabled by the application of labelling chemistries. This dissertation presents novel analytical workflows and data analysis routines to overcome current challenges in proteomics methods for the identification of protein-protein interactions (PPIs) and the study of protein conformation and dynamics. Affinity purification followed by mass spectrometry (AP-MS) is a prominent approach in the study of PPIs. However, the conventional workflow suffers from low enrichment efficiencies. I present and evaluate a fluidic platform that captures and processes ultralow nanoliter quantities of magnetic particles, simultaneously increasing the efficiency of PPI detection and strongly suppressing non-specific binding. It enables the study of protein conformational analysis directly from cells as I demonstrate first by describing new concepts in data analysis for hydrogen/deuterium exchange mass spectrometry (HX-MS) and second by applying them to proteins isolated directly from cells.
dc.identifier.citation	Raval, S. (2023). Enabling structural proteomics with high efficiency protein enrichment technology (Doctoral thesis, University of Calgary, Calgary, Canada). Retrieved from https://prism.ucalgary.ca.
dc.identifier.uri	https://hdl.handle.net/1880/117130
dc.identifier.uri	https://doi.org/10.11575/PRISM/41972
dc.language.iso	en
dc.publisher.faculty	Graduate Studies
dc.publisher.institution	University of Calgary
dc.rights	University of Calgary graduate students retain copyright ownership and moral rights for their thesis. You may use this material in any way that is permitted by the Copyright Act or through licensing that has been assigned to the document. For uses that are not allowable under copyright legislation or licensing, you are required to seek permission.
dc.subject	Affinity Purification
dc.subject	Mass Spectrometry
dc.subject	Hydrogen-Deuterium Exchange
dc.subject	Bioanalytical Chemistry
dc.subject.classification	Chemistry--Analytical
dc.subject.classification	Biochemistry
dc.title	Enabling Structural Proteomics with High Efficiency Protein Enrichment Technology
dc.type	doctoral thesis
thesis.degree.discipline	Chemistry
thesis.degree.grantor	University of Calgary
thesis.degree.name	Doctor of Philosophy (PhD)
ucalgary.thesis.accesssetbystudent	I do not require a thesis withhold – my thesis will have open access and can be viewed and downloaded publicly as soon as possible.