A Semi-Synthetic Method for Sythesis of SUMOylated Hypoxia Inducible Factor-1 alpha for Future Functional Analysis

Abstract: Native chemical ligation (NCL) is a method used to synthesize a covalent bond between two proteins. The method has been extensively used to react a C-terminal thioester on one protein with an N-terminal cysteine residue on another protein. Our strategy deviates from this usual method by incorporating a cysteine containing analogue of pyrrolysine (PCA) into any site on a recombinant protein. My work uses NCL to conjugate Small Ubiquitin like Modifier (SUMO) to Hypoxia Inducible Factor -1α (HIF-1α) prelabeled with PCA. HIF-1α is a transcription factor that is known to generate a protective response during hypoxia, a common symptom of stroke in which there are depleted oxygen levels in tissues. Furthermore, during hypoxia, SUMO is conjugated to HIF-1α at Lys391 and Lys477. An in vitro preparation of SUMOylated HIF-1α is desired because of two challenges in isolating the protein from in vivo preparations; first, HIF-1α is rapidly degraded in normal respiring tissues and second, deconjugating enzymes cleave SUMO away once the function of the SUMOylated form is mediated in the cell. In order to generate the in vitro conjugated product we use the principles of intein chemistry to generate a thioester on SUMO. We incorporate PCA at previously predicted SUMOylation sites and set up a reaction with the SUMO derived thioester. We purified the conjugated products and this conjugated product will be utilized to create better anti-ischemia agents for patients with strokes or metabolic disorders that require high oxygen demands.Our research is supported by a grant from the American Heart Association (10POST3900010) awarded to M.S.NA five-year embargo was granted for this item

Do the Prophets Teach That Babylonia Will Be Rebuilt in the Eschaton

Dispensationalists have traditionally argued that Babylon in Revelation 14 and chaps. 17-18 is a symbol indicating some form of reestablished Rome. * In recent days a renewed interest has been shown in the idea that the ancient empire of Babylonia and city of Babylon will be rebuilt.2 This conclusion comes from a reading of the prophets—Isaiah and Jeremiah in particular—in a manner that requires the rebuilding of the city and empire of Babylonia in the eschaton. My approach to this question is from three different perspectives: (1) to study the context of the oracles against the nations (OAN) found in Isaiah 13-23 and, in particular, the way the critical thirteenth chapter fits into the Sitz im Leben of the eighth century during which Isaiah was prophesying; (2) to study the language of destruction found in this same unit and relate it to the treaty curses found in the ancient Near East and to the rest of the OT; and (3) to examine Jeremiah\u27s prophecies against Babylon containing much of the same language as that of Isaiah

Social Media is Bananas: The effect social media has on rebranding and customer attraction and retention

The primary purpose of this study was to investigate whether social media had an impact on the rebranding of the Savannah Bananas baseball team. The study also focuses on the impact social media has had on the attraction and retention of customers. Interviews were conducted with full time staff members and customers of the Savannah Bananas. An online survey was distributed through social media to learn more about the connection between customers and the Bananas social media. The results of the study suggest that social media has had a positive affect on both the rebranding of the Savannah Bananas and the attraction and retention of customers