Note: This work of creative scholarship is rooted in collaboration between three female artist-scholars: Carly Bates, Raji Ganesan, and Allyson Yoder. Working from a common intersectional, feminist framework, we served as artistic co-directors of each other’s solo pieces and co-producers of Negotiations, in which we share these pieces alongside each other. Negotiations is not a showcase of three individual works, but a conversation among three voices. As collaborators, we have been uncompromising in the pursuit of our own unique inquiries and voices and each of our works of creative scholarship stand alone. However, we believe that all of the parts are best understood in relationship to each other and to the whole. For this reason, we have chosen to cross-reference our thesis documents here, and we encourage readers to view the performance of Negotiations in its entirety.
Thesis documents cross-referenced:
French Vanilla: An Exploration of Biracial Identity Through Narrative Performance, by Carly Bates
Bhairavi: A Performance-Investigation of Belonging and Dis-Belonging in Diaspora Communities, by Raji Ganesan
Deep roots, shared fruits: Emergent creative process and the ecology of solo performance through “Dress in Something Plain and Dark,” by Allyson Yoder
The membrane proximal region (MPR, residues 649–683) and transmembrane domain (TMD, residues 684–705) of the gp41 subunit of HIV-1’s envelope protein are highly conserved and are important in viral mucosal transmission, virus attachment and membrane fusion with target cells. Several structures of the trimeric membrane proximal external region (residues 662–683) of MPR have been reported at the atomic level; however, the atomic structure of the TMD still remains unknown. To elucidate the structure of both MPR and TMD, we expressed the region spanning both domains, MPR-TM (residues 649–705), in Escherichia coli as a fusion protein with maltose binding protein (MBP). MPR-TM was initially fused to the C-terminus of MBP via a 42 aa-long linker containing a TEV protease recognition site (MBP-linker-MPR-TM).
Biophysical characterization indicated that the purified MBP-linker-MPR-TM protein was a monodisperse and stable candidate for crystallization. However, crystals of the MBP-linker-MPR-TM protein could not be obtained in extensive crystallization screens. It is possible that the 42 residue-long linker between MBP and MPR-TM was interfering with crystal formation. To test this hypothesis, the 42 residue-long linker was replaced with three alanine residues. The fusion protein, MBP-AAA-MPR-TM, was similarly purified and characterized. Significantly, both the MBP-linker-MPR-TM and MBP-AAA-MPR-TM proteins strongly interacted with broadly neutralizing monoclonal antibodies 2F5 and 4E10. With epitopes accessible to the broadly neutralizing antibodies, these MBP/MPR-TM recombinant proteins may be in immunologically relevant conformations that mimic a pre-hairpin intermediate of gp41.