Spider silk is a novel material with high strength and toughness and is lightweight, making it attractive for high-performance fiber and biomedical applications. The amino acid sequence and molecular architecture of spider silk protein are becoming known gradually with science and technology development. However, its relationship of structure and function is a puzzle which attracts materials scientists to unveil it. In this study we investigated the changes of the crystal structure as a result of different processing techniques to spider silk protein. Although the crystallization of biomacromolecules is very difficult and complex, we did observe the crystal of electrospun spider silk protein treated by external high pressure in its solid state. This study also provides us a simple and effective method to prepare protein crystals. It is most important that this is the first time that we found spider silk protein as a result of electrospinning that had a crystallization memory property when it was stimulated by a high pressure, which will further extend the list of its particular properties. This finding will also lead us to study it deeply and widen our research on proteins of other organisms.