DsRed-Monomer as bifunctional tag

ANYL 122

Ann M Goulding, agouldin@iupui.edu1, Suresh Shrestha2, Yasmeen Rahimi3, Eric Hunt2, and Sapna K Deo1. (1) Department of Chemistry and Chemical Biology, Indiana University Purdue University Indianapolis, 402 N Blackford St, LD 326, Indianapollis, IN 46202, (2) Dept of Chemistry and Chemical Biology, Indiana University Purdue University Indianapolis, 402 N Blackford St, Indianapolis, IN 46202, (3) Department of Chemistry & Chemical Biology, Indiana University Purdue University Indianapolis, 402 N. Blackford Street, LD326, Indianapolis, IN 46202
Recent work in our laboratory demonstrated that a purification of the red fluorescent protein, DsRed-monomer, can be performed using immobilized copper. Now we have performed studies to demonstrate application of DsRed-monomer as an affinity tag for the purification of peptides/proteins of interest fused to the DsRed-monomer. The work performed toward purification of DsRed-monomer-bradykinin, and the subsequent development of assay for bradykinin using DsRed-monomer as a fluorescence tag will be discussed. In addition, application of DsRed-monomer in the isolation of interacting protein was evaluated. For this study, a model bait–ligand pair, namely M13-calmodulin (CaM) was selected. We have constructed a fusion between DsRed-monomer and a CaM-binding peptide (M13) as the bait peptide to fish out CaM from the cellular matrix. The results of this study in the extraction of CaM from the cellular matrix using DsRed-monomer-M13 fusion protein will be presented.
 

General Posters
7:00 PM-9:00 PM, Sunday, August 19, 2007 BCEC -- Exhibit Hall - B2, Poster

Division of Analytical Chemistry

The 234th ACS National Meeting, Boston, MA, August 19-23, 2007