Purification and Crystallization of Bacterial Direct Oxygen Sensor Phosphodiesterase PAS Domain
Session Number
A02
Advisor(s)
Xiaojing Yang, University of Illinois at Chicago
Xiaoli Zeng, University of Illinois at Chicago
Location
B-110
Start Date
28-4-2016 10:15 AM
End Date
28-4-2016 10:35 AM
Abstract
All living organisms have biological sensors that receive signals and regulate cell functions in the presence of oxygen. One of these particular sensor proteins, Direct Oxygen Sensor Phosphodiesterase (DosP) from E.coli, works with a partner protein to regulate the presence of cyclic-di-GMP, an important dinucleotide second messenger. In this investigation, the oxygen sensing module of DosP, referred to as DosP 382, was examined by means of protein purification and X-ray crystallography. Purification experiments were conducted by amplification of the sequence of DosP 382, insertion into an expression vector, transformation into an E.coli culture, extraction and elution. Crystallization experiments were set up with purified DosP 382 and observed under a microscope. Preliminary results have shown successful protein purification, however the outcomes from crystallization experiments have failed to produce any protein crystals. Further investigation of DosP 382 may reveal greater intricacies of its conformational changes in response to oxygen sensing, as well as a better understanding of certain bacterial cell processes.
Purification and Crystallization of Bacterial Direct Oxygen Sensor Phosphodiesterase PAS Domain
B-110
All living organisms have biological sensors that receive signals and regulate cell functions in the presence of oxygen. One of these particular sensor proteins, Direct Oxygen Sensor Phosphodiesterase (DosP) from E.coli, works with a partner protein to regulate the presence of cyclic-di-GMP, an important dinucleotide second messenger. In this investigation, the oxygen sensing module of DosP, referred to as DosP 382, was examined by means of protein purification and X-ray crystallography. Purification experiments were conducted by amplification of the sequence of DosP 382, insertion into an expression vector, transformation into an E.coli culture, extraction and elution. Crystallization experiments were set up with purified DosP 382 and observed under a microscope. Preliminary results have shown successful protein purification, however the outcomes from crystallization experiments have failed to produce any protein crystals. Further investigation of DosP 382 may reveal greater intricacies of its conformational changes in response to oxygen sensing, as well as a better understanding of certain bacterial cell processes.