Expression, Purification and Crystallization of Heat Shock Factor Binding Protein 1*

Abstract

Heat shock response rises when the organisms in the environmental stimulate. During heat shock, a lot of proteins which is so called heat shock protein that do not express in normal state largely express. These proteins′ expression is regulated by a family of transcription factor. These proteins were called heat shock transcriptional factors (HSFs). Among these HSFs, HSF1 is the most important protein. And HSFs are also regulated by other proteins, such as heat shock factor binding protein1 (HSBP1). HSBP1 interact with the trimer formation of HSF1 to convert the HSF1 from its active trimer state to inert monomer state. In order to do the further functional investigation, the gene of HSBP1 was cloned by PCR using the virus cDNAs as templates and expressed in Escherichia coli BL21 (DE3). Furthermore, the expressed HSBP1 protein was purified and crystallized. The distinct crystal form was obtained by the hanging-drop vapor-diffusion process to carry on screening and the optimization to the crystallization condition. The crystals belong to R3 space group with the cell parameter a = b = 35.2, c = 233.3.