Characterizing fine protein structure of Rubredoxin variants across Pyrococcus and Polaromonas: A comparative analysis

Location

Suwanee, GA

Start Date

7-5-2024 1:00 PM

End Date

7-5-2024 4:00 PM

Description

Introduction: Extremophiles are a group of organisms defined by their ability to sustain life in Earth’s most uninhabitable conditions. These organisms have sophisticated machinery to tolerate unforgiving pressures, pH, and temperatures– high and low. They span all three domains– Archaea, Prokarya, and Eukarya. Notably, a species of archaea called Pyrococcus furiosus, was one of the first hyperthermophiles to be examined. P. furiosus studies shaped archaeon studies for later discovered members of its genus and other extremophiles.

Many bacteria and archaea express a relatively small, iron-binding protein called rubredoxin. Rubredoxins are active agents in redox metabolism, thanks in part to their coordination chemistry. An iron-cysteine cluster is found at its core in a stable, tetrahedral geometry, which facilitates electron transfer in metabolic pathways. Rubredoxin domains also serve as integral primary and secondary metal binding sites for larger metalloproteins.

Methods: For our characterization studies, we isolated and purified rubredoxin from three extremophiles– Pyrococcus furiosus, as a representative hyperthermophile, Polaromonas glacialis (a psychrophile), and Pyrococcus yayanosii (a piezophile). Electrospray ionization mass spectrometry (ESI-MS) and inductively coupled plasma mass spectrometry (ICP-MS) analyses were employed to contrast potential differences in molecular weight and metal composition, respectively. At this time, characterization measures include ultraviolet-visible spectra analysis. Additional characterization techniques will be pursued in the future.

Results: Analysis of results is ongoing. These studies will aid us as we further contrast the amino acid residues which are adapted for each organism’s unique metabolic requirements and growth conditions.

Discussion: Rubredoxins are proteins distinct in their ability to house and coordinate metal ions. Characterization studies are the first step of understanding rubredoxin’s structure and behavior in-vivo. We hope that these insights will guide our future studies on rubredoxin’s fine structure and optimization for biomedical application.

Embargo Period

5-23-2024

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COinS
 
May 7th, 1:00 PM May 7th, 4:00 PM

Characterizing fine protein structure of Rubredoxin variants across Pyrococcus and Polaromonas: A comparative analysis

Suwanee, GA

Introduction: Extremophiles are a group of organisms defined by their ability to sustain life in Earth’s most uninhabitable conditions. These organisms have sophisticated machinery to tolerate unforgiving pressures, pH, and temperatures– high and low. They span all three domains– Archaea, Prokarya, and Eukarya. Notably, a species of archaea called Pyrococcus furiosus, was one of the first hyperthermophiles to be examined. P. furiosus studies shaped archaeon studies for later discovered members of its genus and other extremophiles.

Many bacteria and archaea express a relatively small, iron-binding protein called rubredoxin. Rubredoxins are active agents in redox metabolism, thanks in part to their coordination chemistry. An iron-cysteine cluster is found at its core in a stable, tetrahedral geometry, which facilitates electron transfer in metabolic pathways. Rubredoxin domains also serve as integral primary and secondary metal binding sites for larger metalloproteins.

Methods: For our characterization studies, we isolated and purified rubredoxin from three extremophiles– Pyrococcus furiosus, as a representative hyperthermophile, Polaromonas glacialis (a psychrophile), and Pyrococcus yayanosii (a piezophile). Electrospray ionization mass spectrometry (ESI-MS) and inductively coupled plasma mass spectrometry (ICP-MS) analyses were employed to contrast potential differences in molecular weight and metal composition, respectively. At this time, characterization measures include ultraviolet-visible spectra analysis. Additional characterization techniques will be pursued in the future.

Results: Analysis of results is ongoing. These studies will aid us as we further contrast the amino acid residues which are adapted for each organism’s unique metabolic requirements and growth conditions.

Discussion: Rubredoxins are proteins distinct in their ability to house and coordinate metal ions. Characterization studies are the first step of understanding rubredoxin’s structure and behavior in-vivo. We hope that these insights will guide our future studies on rubredoxin’s fine structure and optimization for biomedical application.