Project Number: 5306-51530-019-77
Project Type: Specific Cooperative Agreement

Start Date: Dec 01, 2012
End Date: Jul 31, 2013

Objective:
Long- and medium-chain acylcarnitines have been shown to be pro-inflammatory and inhibit insulin-signaling at p-Akt. Crucial to understanding this biology, and the biology of other lipid moieties that are thought to impinge upon muscle insulin signaling, is to ask "Are these metabolites trafficked or sequestered in muscle, might such dynamics affect their bioactivities, and would the trafficking be regulated under altered metabolic states?"

Approach:
In collaboration with the Randen Patterson lab (UC Davis), the hypothesis that myoglobin is a heretofore unappreciated site of control for regulation of free fatty acid levels and acylcarnitines in muscle will be tested. If true, this could impinge upon insulin action and muscle metabolism generally. We anticipate that the experiments to test this postulate will take about 6 months to perform, and will include: (a) computer modeling of various chain lengths of acylcarnitines binding to myoglobin, including the effects of pH, oxygen tension, and other relevant biochemical conditions; (b) cell culture and biochemical assays to confirm the in silico models—specifically, labeled palmitoylcarnitine and other relevant chain-lengths of acylcarnitines will be used in cell models used routinely in the Patterson lab to test for association/binding with overexpressed myoglobin, with appropriate negative controls, etc. Additional possible studies include metabolomics analyses of isolated myoglobin protein to ascertain if acylcarnitines and fatty acids are bound to myoglobin tightly.