Protein Structure Analysis

Primary Structure

It’s the most basic level, representing the linear order of amino acids in a polypeptide chain.

Secondary Structure

It includes regular structural motifs such as spirals and pleated formations that arise early in the folding process.

Tertiary Structure

It results from the full folding of coils and sheets, driven by various forces like hydrogen bonding, ionic interactions, and disulfide linkages between side chains.

Quaternary Structure

It’s the highest level of organization, formed when several individual chains interact and stabilize each other to create the final protein.

Aim of Protein Structure Lab

Deepen understanding of biologically active protein molecules

Importance of protein electrophoresis

What are the two types of gel systems?

Denaturing and non denaturing

Non-denaturing gel system

Protein separation that preserves protein function and activity

Denaturing gel system

Denatures the protein into subunit to exam the structure

What does SDS-PAGE stand for?

Sodium dodecyl sulfate polyacrylamide gel electrophoresis

What type of gel system is SDS-PAGE?

Denaturing

What is a reducing agent used for in the protein structure analysis lab?

Breaks disulfide bonds

Principles of the protein structure analysis lab

Compare reduced samples to unreduced samples to determine presence of disulfide bonds

Reagents used in Protein Structure Lab

3 protein samples

TCEP reducing agent

How is SDS-PAGE different from agarose gel electrophoresis?

SDS-PAGE separates samples by molecular weight, but agarose GE separates samples by size