The use of the Knob-Socket model in the synthesis and expression of a novel protein, Star1.0

Poster Number

11C

Lead Author Major

Biohemistry

Lead Author Status

Sophomore

Second Author Major

Biochemistry

Second Author Status

Junior

Format

Poster Presentation

Faculty Mentor Name

Jerry Tsai

Faculty Mentor Department

Chemistry

Graduate Student Mentor Name

Taylor Crawford

Graduate Student Mentor Department

Chemistry

Abstract/Artist Statement

The knob-socket model is a code to describe how proteins interact to form tertiary structures. The basis of the knob-socket model includes a one amino acid residue knob from one piece of secondary structure packing into a three amino acid residue socket from another piece of secondary structure. Sockets can be described as free (disfavoring knob packing), or filled (favoring knob packing) and are given these designations depending on their three amino acid composition. A propensity library was developed and gives the frequency at which each socket is found to be either free or filled based on data from the PDB database. We aim to show that the knob socket model and the socket propensity library can be used to predict protein secondary structure, and therefore be used in protein design. This idea was used in the de novo design of the STAR1.0 protein. The STAR1.0 protein was designed with a unique five alpha-helical structure in the shape of a five pointed star. The sequence was developed using the alpha-helical propensity library and was further optimized for expression in E. coli. The STAR1.0 protein was expressed and purified, and then secondary structure was analyzed using circular dichroism spectroscopy. STAR1.0 is now being expressed on a large scale and purified in order to produce a high concentration of protein for x-ray crystallography, which will determine if the structure matches the theoretical five pointed star shape.

Location

DeRosa University Center, Ballroom

Start Date

29-4-2017 1:00 PM

End Date

29-4-2017 3:00 PM

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Apr 29th, 1:00 PM Apr 29th, 3:00 PM

The use of the Knob-Socket model in the synthesis and expression of a novel protein, Star1.0

DeRosa University Center, Ballroom

The knob-socket model is a code to describe how proteins interact to form tertiary structures. The basis of the knob-socket model includes a one amino acid residue knob from one piece of secondary structure packing into a three amino acid residue socket from another piece of secondary structure. Sockets can be described as free (disfavoring knob packing), or filled (favoring knob packing) and are given these designations depending on their three amino acid composition. A propensity library was developed and gives the frequency at which each socket is found to be either free or filled based on data from the PDB database. We aim to show that the knob socket model and the socket propensity library can be used to predict protein secondary structure, and therefore be used in protein design. This idea was used in the de novo design of the STAR1.0 protein. The STAR1.0 protein was designed with a unique five alpha-helical structure in the shape of a five pointed star. The sequence was developed using the alpha-helical propensity library and was further optimized for expression in E. coli. The STAR1.0 protein was expressed and purified, and then secondary structure was analyzed using circular dichroism spectroscopy. STAR1.0 is now being expressed on a large scale and purified in order to produce a high concentration of protein for x-ray crystallography, which will determine if the structure matches the theoretical five pointed star shape.