ABSTRACT

The Illinois Junior Academy of Science

This form/paper may not be taken without IJAS authorization.

CATEGORY Biochemistry STATE REGION # 6

SCHOOL Niles North High School IJAS SCHOOL # 6038
CITY/ZIP Skokie, IL 60077 SCHOOL PHONE # 847.626.2254
SPONSOR Christi Camel

MARK ONE: EXPERIMENTAL INVESTIGATION DESIGN INVESTIGATION

NAME OF SCIENTIST* Sumona Banerjee GRADE 12

NAME OF SCIENTIST GRADE
NAME OF SCIENTIST GRADE
NAME OF SCIENTIST GRADE

* If this project is awarded a monetary prize, the check will be written in this scientist's name, and it will be his/her responsibility to distribute the
prize money equally among all participating scientists.

PROJECT TITLE The Effect of Mutations in the Ligand-Binding Pocket on the Function of Ftz-F1

Purpose: The purpose of this study is to create mutations of orphan receptor Ftz-F1 by changing the dynamics of its ligand binding
pocket through mutagenesis and determine how do these mutations affect the stability of Ftz F1 and its binding ability to its coactivator
Ftz.We hypothesized that The G913K + peptide, G913Y + peptide, and L907F + peptide samples should be more stable than the wild
type + peptide sample. This is because the increase in amino acid mass should decrease the speculated movement of helix 6. L907V +
peptide samples should be less stable than the wild type + protein sample due to the opposite reasoning.

Procedure: To conduct this experiment, PCR mutagenesis was used to introduce mutations in the Ftz F1 ligand binding domain. These
mutations were sequenced and then transformed into bacteria. Afterwards, the bacteria was lysed and purified for protein samples. A
thermal shift assay was used to determine the melting point of each protein sample as well as the melting points of each protein while
being bound to the LxxLL peptide.

Conclusion: Two of the three hypotheses were supported. The melting points of GK + peptide and GY + peptide were significantly
higher than the wild type + peptide melting point, thus exhibiting a higher stability. The LF + peptide and LV + peptide samples
displayed displayed the opposite. The applications of these findings relate to the engineering of synthetic ligands as a means of drug
discovery. A decrease in amino acid size may be correlated to a decrease in protein stability and thus the down regulation of the
appropriate gene. This may be applied to various human diseases activated by nuclear receptors such as cancer, diabetes, arthritis etc.

1) Limit Abstract to 3 paragraphs (about 200 words or less). a) Purpose - what you set out to investigate; b) Procedure - how you did
it; c) Conclusion - based on your results. Label each paragraph.
2) Must be typed, single-spaced on the front of this form. Do not write on the back of this form.
3) Three copies of your complete paper are required at the State Science Project Exposition.
Four copies of your complete paper are required for the State Paper Session Competition.

This form must be used. This form must be displayed on the front of the exhibitors display board. It may be reduced to half a sheet of paper;
8.5 inches (vertical) X 5.5 inches (horizontal).