Student Theses and Dissertations

Date of Award

2012

Document Type

Thesis

RU Laboratory

Hang Laboratory

Abstract

Lipidation of proteins regulates many cellular processes such as signaling transduction and membrane sorting by modulating protein localization and proteinprotein interactions. As such, defects in protein lipidation can render host cells more susceptible to microbial infection and are also associated with a variety of human diseases ranging from cancer to neurological disorders. Moreover, viral and bacterial pathogens can exploit and modulate the host lipidation machinery to enhance infection. Robust biochemical methods for characterizing lipidated proteins are therefore important for understanding fundamental physiology and disease mechanisms. In this thesis, I report the development of alkyne-lipid chemical reporters that afford more sensitive detection and proteomic analysis of fatty-acylated and prenylated proteins using bioorthogonal ligation methods. Alkynyl-fatty acids of 14 and 18 carbons afforded enhanced detection of N-myristoylated and S-palmitoylated proteins, respectively, while alkynyl-farnesol allowed more robust analysis of S-prenylated proteins. These new lipid chemical reporters enabled the discovery of bacterial virulence factors that are lipidmodified by host enzymes during infection as well as lipidated host proteins (IFITM3 and ZAP) involved in cellular resistance to viruses. My thesis work highlights the utility of alkyne lipid reporters for biochemical analysis of lipidated proteins at the host-pathogen interface that will hopefully help other biological studies.

Comments

A thesis presented to the faculty of The Rockefeller University in partial fulfillment of the requirements for the degree of Doctor of Philosophy.

Permanent URL

http://hdl.handle.net/10209/520

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