Micrometer and submicrometer diameter fibers of recombinant dragline spider silk analogues, synthesized via protein engineering strategies, have been electrospun from 1, 1, 1, 3,3,3-hexafluoro-2-propanol (HFIP) and compared with cast films via Raman spectroscopy in order to assess changes in protein conformation that may result from the electrospinning process. Although the solvent casting process was shown to result in predominantly β-sheet conformation similar to that observed in the bulk, the electrospinning process causes a major change in conformation from β-sheet to α-helix. A possible mechanism involving electric field-induced stabilization of a-helical segments in HFIP solution during the electrospinning process is discussed.