灵芝免疫调节蛋白LZ-8的制备和晶体分析

LZ-8蛋白是从灵芝菌丝中分离到的真菌免疫调节蛋白,具有多种免疫调节功能,然而这一蛋白的作用机制尚不清楚。通过蛋白质晶体结构的解析,能够得到蛋白质空间结构特点,从而阐述蛋白质功能的机制。旨在得到LZ-8蛋白的晶体,并获得空间结构数据。以pET21a为表达载体,获得诱导表达的rLZ-8,通过亲和层析、分子筛凝胶层析和阴离子交换层析纯化,蛋白纯度在98%以上,采用悬滴气相扩散法得到蛋白晶体,并获得3.2?数据,为进一步对真菌免疫调节蛋白功能和结构的研究奠定了基础。

Expression and crystallographic studies of a fungal immunomodulatory protein LZ-8 from a medicinal fungus Ganoderma lucidum

LZ-8 protein, isolated from a well known Chinese traditional medicinal fungus Ganoderma lucidum, is the first member of fungal immunomodulatory protein, members of which have been isolated from a variety of medical and edible mushrooms in the last two decades. The protein plays a multifunctional and important role in modulating immune system. In this report, in order to get LZ-8 protein crystals, the LZ-8 gene was expressed and purified by affinity chromatography, gel filtration and iron exchange subsequently. The protein was then crystallized using the hanging-drop vapour-diffusion method. The LZ-8 crystals were obtained and the phase information was calculated by X-ray diffraction. The resolution of LZ-8 crystals is 3.2?. This study will provide an insight into the structure of fungal immunomodulatory proteins.