VCU Bioinformatics and Bioengineering Summer Institute
Virginia Commonwealth University
imageimageHomeBio What?The InstituteThe People
The Institute
Goals of the Institute
Two-year Plan
Course web pages
News
Archives
Application process
About the BBSI

Research Simulation Scenario
Protein Overexpression
Exploring Mutant Proteins

Analysis of mutations
You sequence all the mutant proteins from the rare fluorescent colonies. This gives you the identities of the amino acid changes that leads to moderate decreases in precipitation. If you could understand why these changes work, then perhaps you could figure out how to make even more effective changes, either in other amino acids or by combining mutations.

Wild-type UDP-glc DH
...SKVDKINNGLSPIQDEYIEY...

Mutant UDP-glc DH
...SKVEKINNGLSPIQDEYIEY...
...SKVDKINNGVSPIQDEYIEY...
...SKVDKINNGLSPIQDESIEY...
You wish you could visualize where on the protein's three dimensional structure the effective mutations are situated, for then you might learn which other amino acids in the vicinity are candidates for directed mutagenesis. This way you might rationally design a protein that won't precipitate.

There IS a three-dimensional structure known for a related protein, UDP glucose dehydrogenase from Streptococcus pyogenes, but nothing structural is known about YOUR protein. Tough luck?

Back to main Scenario page      back one page     continue