PP1 Inhibitor 2 in Zebrafish

Poster Number

51

Lead Author Major

Pre-Dentistry

Format

Poster Presentation

Faculty Mentor Name

Doug Weiser

Faculty Mentor Department

Biological Sciences

Abstract/Artist Statement

Protein phosphatase 1 (PP1) is essential regulator of the control of glycogen metabolism, muscle contraction, cell division and regulation of membrane receptors and channels. Regulation of these processes is through PP1 holoenzymes that facilitates the PP1 catalytic subunit. These holoenzymes consists, of different regulatory and inhibitory subunits along with the catalytic subunit. Protein phosphatase inhibitor-2 (I-2) is one of the earliest discovered inhibitors, but still much is not known about it and it’s role in vivo. The main topic of research is how it binds to certain PP1 subunits and not others. A goal of our project is to examine and potentially develop zebrafish I-2 as a genetic model system. Inhibition by I-2 is used to distinguish PP1 activity from other phosphatases. Due to lack of research on this inhibitor we are running controls in order to purify the protein and then run experiments on the same inhibitor in zebrafish. We are currently trying to see the differential interactions of I-2 with various PP1 holoenzymes. Recombinant human I-2 was purified from E. Coli. A pull down with GST I-2 and GST was done against a series of PP1 regulatory subunits: Gadd34, GaddRVXF (non PP1 binding mutant), Flag-Neurabin, Flag-Crep and MYPT1. This GST-pull down will be able to distinguish with ones bind and which do not, which will allow us to examine the evolutionary conservation of I-2 function.

Location

DeRosa University Center, Ballroom

Start Date

20-4-2013 1:00 PM

End Date

20-4-2013 3:00 PM

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Apr 20th, 1:00 PM Apr 20th, 3:00 PM

PP1 Inhibitor 2 in Zebrafish

DeRosa University Center, Ballroom

Protein phosphatase 1 (PP1) is essential regulator of the control of glycogen metabolism, muscle contraction, cell division and regulation of membrane receptors and channels. Regulation of these processes is through PP1 holoenzymes that facilitates the PP1 catalytic subunit. These holoenzymes consists, of different regulatory and inhibitory subunits along with the catalytic subunit. Protein phosphatase inhibitor-2 (I-2) is one of the earliest discovered inhibitors, but still much is not known about it and it’s role in vivo. The main topic of research is how it binds to certain PP1 subunits and not others. A goal of our project is to examine and potentially develop zebrafish I-2 as a genetic model system. Inhibition by I-2 is used to distinguish PP1 activity from other phosphatases. Due to lack of research on this inhibitor we are running controls in order to purify the protein and then run experiments on the same inhibitor in zebrafish. We are currently trying to see the differential interactions of I-2 with various PP1 holoenzymes. Recombinant human I-2 was purified from E. Coli. A pull down with GST I-2 and GST was done against a series of PP1 regulatory subunits: Gadd34, GaddRVXF (non PP1 binding mutant), Flag-Neurabin, Flag-Crep and MYPT1. This GST-pull down will be able to distinguish with ones bind and which do not, which will allow us to examine the evolutionary conservation of I-2 function.