Analyze the role of HAI-2 N-Glycosylation in matriptase activation mechanism

Ru Jia (Mentor: Dr. Chen-Yong Lin, Lombardi Comprehensive Cancer Center, Georgetown University)

December 4th, 2015, 2:00pm-2:30pm, Room 1202, Harris Building

Matriptase is an epithelial-derived, integral membrane serine protease, which is synthesized in zymogen form, and to work with active as an enzyme. It plays a very important role in breast cancer invasion and metastasis. According to previous research, the activation of matriptase must need HAI-1 expression. But now evidence show that some cancer cells still have high matriptase activity without HAI-1 expression, which means there might exist other inhibitors that may replace the activation role of HAI-1. After blast HAI-1 and other Kunitz type serine protease inhibitor, I find that HAI-2 may have the similar ability to bind with matriptase as HAI-1.Therefore, the main focus of my project is aim to verify if this hypothesis is validate. First we annotate HAI-2, predict its action sites, and get the Western Blot results as foundations. Next, we filter 5 N-linked glycosylation enzymes from related database, in order to analyze and pick primers for the following RT-PCR step. Also, according to Ying-Jung J. Lai [1], antibody XY9 and DC16 were chosen in sequence screening.

[1] Ying-Jung J. Lai et al, “N-Glycan Branching Affects the Subcellular Distribution of and Inhibition of Matriptase by HAI-2/Placental Bikunin”, PLOS ONE. 2015 July; 3(7): 1-16.