NMNAT1, A Protein Linked With Blinding Diseases Is Post-Translationally Lipid Modified By Protein Palmitoylation

Anthony Siler*, David Sokolov, and Saravanan Kolandaivelu
Departments of Ophthalmology and Visual Sciences, and Biochemistry, West Virginia University Eye Institute, One Medical Center Drive, Morgantown, WV-26506

Presentation No.: 11

Assigned Category (Presentation Format): Biological & Health Sciences (Oral Presentations)

Student’s Major: Biology

Nicotinamide mononucleotide adenylyl transferase-1 (NMNAT1), a protein exclusively present in the nucleus responsible for NAD+ synthesis, plays a vital role in metabolism and redox biology. Even though NMNAT1 is ubiquitously expressed, mutations can lead to severe blinding diseases of which the precise mechanisms are not yet fully understood. This is the first study demonstrating that NMNAT1 is post translational lipid modified by “palmitoylation”. In general, palmitoylation plays diverse roles in protein sorting, localization, stability, activity, and subcellular trafficking. To understand further, we focused to identify the DHHC-domain containing acyl transferase that is responsible for NMNAT1 palmitoylation. To address this, we will co-transfect NMNAT1 construct with DHHC enzymes into HEK-293T cells. Furthermore, acyl-resin assisted capture (Acyl-RAC) method was performed to evaluate NMNAT1 palmitoylation. To determine the palmitoylation role in protein stability and subcellular localization, we will inhibit NMNAT1 palmitoylation with palmitoyl inhibitor 2-Bromopalmitate (2-BP). Overall, our study is crucial for confirming NMNAT1 palmitoylation and determining DHHC enzymes are responsible for palmitoylation, which are important steps toward better understanding NMNAT1 and its role in normal cellular function.

Funding: Saravanan Kolandaivelu Startup Funding

Program/mechanism supporting research/creative efforts: the WVU Vision Research Fellowship Program (Jianha Du)