Defense Date

10-24-2016

Availability

One-year Embargo

Submission Type

dissertation

Degree Name

PhD

Department

Chemistry and Biochemistry

School

Bayer School of Natural and Environmental Sciences

Committee Chair

Partha Basu

Committee Member

Michael Cascio

Committee Member

John F. Stolz

Committee Member

Michael Van Stipdonk

Committee Member

Jesus Tejero Bravo

Keywords

mARC, Mass spectrometry, Molybdenum enzymes, Protein-protein interactions

Abstract

Newly discovered pterin-molybdoenzymes mammalian proteins, mARC1, and mARC2, are though to activate pro-drug, metabolize mutated base pairs, and producing nitric oxide from nitrite. However, the physiological function of the mARC proteins remain unclear. It is hypothesized that identifying partner proteins through protein-protein interactions can provide evidence to their physiological function. This study aimed to further investigate the potential function of mARC proteins through the identification of protein-protein interaction in human cell lysate, utilizing co-immunoprecipitation, crosslinking reagent, and pull-down assays. Several proteins (e.g. mitogen-activated protein kinase 5, phosphatidylinositol 4-phospahte 3-kinase C2 domain-containing subunit beta, ADP/ATP translocase 1, and myotubularin) were identified as interacting partners of mARC1, and mARC2. These interacting proteins are affiliated with various KEGG signaling pathways. From the interacting proteins we infer, mARC1 and mARC2 may be involved in the regulation of nitric oxide signaling under hypoxic. Reverse co-immunoprecipitation experiments using select proteins from the pathways detected mARC proteins. The investigation was then expanded to include pig liver mitochondrial fraction and rat liver cell lysate, in which all experimental conditions identified mARC proteins using reverse co-immunoprecipitation.

Format

PDF

Language

English

Available for download on Wednesday, December 20, 2017

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