Karatan Lab

Alex Rutkovsky
Nick Walter

Home

Teaching
Research

Ece Karatan
[email protected]

Appalachian State University
Department of Biology
Rankin Science North
319 (Office), 303 (Lab)
828-262-6742

Disclaimer

Elucidating the role of polyamine synthesis and transport on
Vibrio cholerae biofilm formation

Our research is involved with quantifying polyamines in different strains of Vibrio cholerae. Our focus is on the polyamine norspermidine, in particular, and how its concentrations inside a biofilm or cell are linked to biofilm formation.

Norspermidine levels have been associated with NspC, NspS, and PotD protein function as part of a mechanism that regulates biofilm development. In order to more deeply understand these mechanisms it is necessary to quantify the amount of norspermidine inside the biofilms made by wildtype and mutant V. cholerae under different environmental conditions. We are achieving this by derivitizing the polyamines from the cell using a benzoylation process, and then quantifying them using HPLC.

Our research is part of the Merck/AAAS Program for undergraduate research, which brings together faculty members and students from Biology and Chemistry departments to work on corss-disciplinary projects. Our research team is composed of Dr. Claudia Cartaya-Marin (Chemistry), Dr. Ece Karatan (Biology), Alex Rutkovsky (a junior representing the Department of Chemistry) and Nick Walter (a junior representing the Department of Biology).

Figure 1. Polyamines putrescine, norspermidine, and spermidine were benzoylated using standard protocols. Benzoylated standards were then separated by HPLC using a Waters Spherisorb ODS2 column and detected by UV-VIS spectroscopy at 229 nm. Various amounts of the standard (green: 12.5 nmoles, yellow: 25 nmoles, blue: 37.5 nmoles, red:50 nmoles) were run in order to construct a calibration curve.