Based on the understanding of different prolamin protein conformations in solution, we presented an assembled nanofabric prepared by incorporating compact zein nanoparticles in electrospun hordein networks. Zein particles in one aspect acted as the plasticizer to decrease the strong hydrogen-bonding interactions among extended hordein molecules. In another, they also played as the reinforcing filler in the flexible hordein matrix. The assembled fibers exhibited significantly improved tensile strength and wet stability in both water and ethanol. The alignment of electrospun fibers further strengthened the nanofabrics in both tangential and normal directions to 17.26 ± 1.41 and 14.02 ± 0.74 MPa, respectively, stronger than that of cancellous bones (5-10 MPa). It has also been discovered that, by simply altering the applied voltage, the resultant hordein/zein nanofabrics can rapidly (within 30 s) form either flat sheets or self-rolled tubes when they were immersed in water. All the fibers demonstrated low toxicity in human primary dermal fibroblast cell culture. Moreover, the electrospun fabrics exhibited a strong resistance to protein adsorption and cell attachment, and the release experiment indicated that they could serve as a carrier for controlled-release of incorporated bioactive compounds into phosphate-buffered saline. Therefore, these electrospun prolamin protein fabrics represent an ideal and novel platform to develop nonadherent drug delivery systems for wound dressing and other biomedical applications.