Screening the effect of bacterially derived peptides on metal homeostasis in mammalian cells

Melhem, Rim 2020, Screening the effect of bacterially derived peptides on metal homeostasis in mammalian cells, B. Science (Hons) thesis, School of Life and Environmental Sciences, Deakin University.

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Title Screening the effect of bacterially derived peptides on metal homeostasis in mammalian cells
Author Melhem, Rim
Institution Deakin University
School School of Life and Environmental Sciences
Faculty Faculty of Science, Engineering and Built Environment
Degree type Honours
Degree name B. Science (Hons)
Thesis advisor Brau, LambertORCID iD for Brau, Lambert
Date submitted 2020-11-06
Keyword(s) Cell culture
Cell bioavailability
Metal chelation
HuTu 80
Summary At the cellular level, trace elements such as iron (Fe), copper (Cu) and zinc (Zn), are required for key cellular functions. Deviations from homeostatic concentrations lead to loss of cellular function and increased disease states. Although they lack efficacy, supplements and probiotics are the most common treatment for metal deficiency. The proposed solution to treat insufficiencies is use bacterially produced pyoverdine siderophores, which have a high affinity towards cations, potentially increasing cellular metal bioavailability. It is hypothesised that pyoverdine increases the biological availability of the metals, in-turn increasing cellular metal levels in a complexed state that prevents the deleterious effects of high intracellular free-metal levels from occurring. Based on Trypan blue exclusion and MTT assays, HuTu 80 cell viability and growth was significantly (p<0.05) increased when exposed to concentrations of up to 0.5 mg/mL for pyoverdine. Cell viability was also assessed for HuTu 80s exposed to varying Fe, Cu, and Zn concentrations, where the addition of pyoverdine produced by Pseudomonas fluorescens DUS1-27 significantly (p<0.05) increased cell viability and growth. Using inductively coupled plasma mass spectrometry, metal levels in HuTu80 cells treated with pyoverdine plus Fe and Cu significantly increased cellular levels by 10-fold and 2-fold, respectively. Gene expression was measured using quantitative real time PCR, with results showing the addition of the chelates and exogenous Fe, Cu and Zn altered the metal homeostasis systems responses. Overall, this study established that pyoverdine increases bioavailability of environmental metal concentrations leading to increased cell viability for mammalian cells.
Language eng
Indigenous content off
Field of Research 1104 Complementary and Alternative Medicine
Description of original 55 p.
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Created: Tue, 15 Dec 2020, 13:23:44 EST by Bernadette Houghton

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