August and the New Semester

Hello Everyone!

It’s good to be back at school! Soon I will be back in the lab getting rolling on a few new ideas.  During the last week of summer research, we discovered that our protein digestion procedure was not optimal for ArsR.  Our proteolytic enzyme, trypsin, which breaks our protein into smaller peptides for bottom-up proteomics does not cleave the protein into small pieces near our residues of interest.  With the electrospray functioning correctly, we were able to see this consistently – we are not getting full coverage of our protein in our spectra because there is a peptide that is too long (larger in mass than our instrument’s capabilities).  We’ve decided to order and try a new enzyme for digestion.

Before we try the new digestion procedure, we’d like to perform cell lysis on an H. pylori culture, digest, and put the entire range of proteins through the HPLC and MS to see if ArsR can be found when it is present at natural levels in an unpurified sample.  Up until now, we’ve been performing tests on a concentrated sample of ArsR.  After this, we will alter digestion and attempt basic purifications before HPLC/MS analysis.

I’m excited to get back down to business!

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