Project Overview: Overexpressing HTT in neuronal cells

My main project revolves around the E3 ligase UBE3A (I will be writing a project overview for this later), but I am also continuing some work in the same vein as Jolene Ho’s work looking at role of HTT in neurological disorders. HTT encodes a protein called huntingtin whose function is not known but appears Read More …

Purification of the HTT C-HEAT domain

I am very excited to write my first post in open notebook. My post on Zenodo summarizes the results of my first ever purification of a huntingtin (HTT) domain. I was able to purify successfully the HTT C-HEAT domain encompassing residues 2088-3144. Figure 1. Anti-FLAG purification of the HTT C-HEAT domain. Eluted protein is seen Read More …

Project Overview

I joined the group of Dr. Cheryl Arrowsmith 2 months ago. Currently, I am working on the Huntington’s disease project along with Dr. Rachel Harding and Jacob McAuley. During these past two months, I have been learning how to purify huntingtin (HTT) protein and overall getting up to date with the project. A brief introduction Read More …

Huntingtin chromatin retention in PARP knock out cells

More forgotten experiments revived by my year-end report. Huntingtin chromatin retention is reduced in PARP KO cells, but in frustration over some confusing results, I overlooked this important data. Lesson: pause and look at the data before rushing on! All the details and links to Zenodo deposits can be found on the Truant lab website.

Purification of putative huntingtin interacting proteins UBE2K, HAP1, NEDD4L

We are getting close to the midpoint of the summer and my project is well underway. With another week passed, I have more purifications to share. Again, I was attempting to purify proteins we think might interact with the huntingtin protein in neuronal cells. This time around was a little less successful than previous posts, Read More …

Production and Purification of Huntingtin interaction partners continued

As the summer continues to progress, so does my project. After the last round of purification, detailed in my previous blog post, I am ready for another round. During this purification, I will be working with SH3G3 and PACN1. PACN1 is responsible for cytoskeletal remodeling through regulation of several other proteins, as well as neuronal Read More …