Title

Determination Of The Phosphorylation Site Of The Regulator Of Iron Transport In Streptococcus Pneumonia By Mass Spectrometry / Liquid Chromatography

Author(s)

Brooks Digh

School Name

South Carolina Governor's School for Science and Mathematics

Grade Level

12th Grade

Presentation Topic

Biochemistry

Presentation Type

Mentored

Mentor

Mentor: Nicholas Grossoehme, Department of Chemistry, Physics, and Geology, Winthrop University

Abstract

Pneumonia is an infection of the lungs that is most commonly caused by the bacteria Streptococcus Pneumonia. Pneumonia kills over one million children under the age of five in the world annually, which is more than any other infectious disease. Pneumonia greatly affects adults over the age of 65, diabetics, heart related illness, smokers and asthmatics. The purpose of this investigation was to determine a secondary cure for Pneumonia that would allow for the better treatment of those who were unable to receive a vaccine. It was hypothesized that the Regulator of Iron Transport (RitR) would be phosphorylated by Serine Threonine Kinase (StkP) somewhere along the DNA binding domain. The two main questions that our research served to answer were: (1) does StkP Phosphorylate RitR, where would the phosphate bonding occur (2) does StkP auto-phosphorylate before phosphorylating RitR? Once the RitR reacted with StkP and was cut into smaller pieces with trypsin digest, mass spectrum analysis was used to determine where RitR was phosphorylated. Through mass spectrometric analysis it was determined that RitR was phosphorylated in a reaction with StkP and ATP. It was also determined that RitR was phosphorylated on the DNA binding domain on the Amino Acid sequence EYDLLATLMGSK. While investigating the reaction of RitR and StkP, it was also discovered that StkP did phosphorylate itself before phosphorylating RitR. Further work will attempt to confirm these results and to test the phosphatase that removes the phosphate form RitR. Then a drug can be developed to take advantage of this weakness and coming ones step closer to finding a cure for this illness.

Start Date

4-11-2015 11:30 AM

End Date

4-11-2015 11:45 AM

COinS
 
Apr 11th, 11:30 AM Apr 11th, 11:45 AM

Determination Of The Phosphorylation Site Of The Regulator Of Iron Transport In Streptococcus Pneumonia By Mass Spectrometry / Liquid Chromatography

Pneumonia is an infection of the lungs that is most commonly caused by the bacteria Streptococcus Pneumonia. Pneumonia kills over one million children under the age of five in the world annually, which is more than any other infectious disease. Pneumonia greatly affects adults over the age of 65, diabetics, heart related illness, smokers and asthmatics. The purpose of this investigation was to determine a secondary cure for Pneumonia that would allow for the better treatment of those who were unable to receive a vaccine. It was hypothesized that the Regulator of Iron Transport (RitR) would be phosphorylated by Serine Threonine Kinase (StkP) somewhere along the DNA binding domain. The two main questions that our research served to answer were: (1) does StkP Phosphorylate RitR, where would the phosphate bonding occur (2) does StkP auto-phosphorylate before phosphorylating RitR? Once the RitR reacted with StkP and was cut into smaller pieces with trypsin digest, mass spectrum analysis was used to determine where RitR was phosphorylated. Through mass spectrometric analysis it was determined that RitR was phosphorylated in a reaction with StkP and ATP. It was also determined that RitR was phosphorylated on the DNA binding domain on the Amino Acid sequence EYDLLATLMGSK. While investigating the reaction of RitR and StkP, it was also discovered that StkP did phosphorylate itself before phosphorylating RitR. Further work will attempt to confirm these results and to test the phosphatase that removes the phosphate form RitR. Then a drug can be developed to take advantage of this weakness and coming ones step closer to finding a cure for this illness.