Submission Title

Crystallization of PEPX from L. helveticus

Presenter Information

Anita Weeks, Whitworth University

Session Number

PS1

Location

Graves Gym

Abstract Number

PS1-p

Abstract

Celiac disease is an autoimmune response triggered by ingestion of gliadin, a glycoprotein present in wheat and barley, which damages the gastrointestinal tract. Enzymatic oral therapy could potentially supplement digestive enzymes so that celiac disease patients could digest gluten without initiating an autoimmune response and damaging their GI tracts. Peptidases of the bacterium Lactobacillus helveticus (L. helveticus) are, theoretically, able to degrade the proline-containing peptides in gluten. The 3D structure of prolyl aminopeptidase (PEPX) from L. helveticus is unknown, therefore characterizing the structure would be beneficial in attempts to design PEPX enzymes with better properties for oral therapy. PEPX was expressed in E. Coli using a pET expression system and purified using nickel affinity chromatography and anion exchange chromatography. Parameters for crystallization of the L. helveticus protein were determined by varying concentrations of PEG and NaCl in phosphate buffer of either pH 5.7 or 6.0.

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Apr 23rd, 10:45 AM Apr 23rd, 12:15 PM

Crystallization of PEPX from L. helveticus

Graves Gym

Celiac disease is an autoimmune response triggered by ingestion of gliadin, a glycoprotein present in wheat and barley, which damages the gastrointestinal tract. Enzymatic oral therapy could potentially supplement digestive enzymes so that celiac disease patients could digest gluten without initiating an autoimmune response and damaging their GI tracts. Peptidases of the bacterium Lactobacillus helveticus (L. helveticus) are, theoretically, able to degrade the proline-containing peptides in gluten. The 3D structure of prolyl aminopeptidase (PEPX) from L. helveticus is unknown, therefore characterizing the structure would be beneficial in attempts to design PEPX enzymes with better properties for oral therapy. PEPX was expressed in E. Coli using a pET expression system and purified using nickel affinity chromatography and anion exchange chromatography. Parameters for crystallization of the L. helveticus protein were determined by varying concentrations of PEG and NaCl in phosphate buffer of either pH 5.7 or 6.0.