Buu A, Garreau H, Jacquet M
Institut de génétique et microbiologie, CNRS URA D1354, Université Paris-XI, Orsay, France.
C R Acad Sci III. 1995 Jun;318(6):665-9.
In order to isolate yeast proteins able to bind to the SH3 domain of the Cdc25p exchange factor, a biochemical approach was used. The SH3 (src homolog type 3) domain of yeast Cdc25p, fused both to a tail of 6 histidine (His) and to glutathione-S-transferase (GST), was purified and then, using His affinity for Ni2+ ions, bound to a Ni-NTA column. This column was used for isolating yeast proteins which have affinity for the yeast SH3-Cdc25p domain. The major protein thus isolated, was sequenced and identified as a yeast glyceraldehyde-3-phosphate dehydrogenase (GAP3DH).
为了分离能够与Cdc25p交换因子的SH3结构域结合的酵母蛋白，采用了一种生化方法。将酵母Cdc25p的SH3（src同源3型）结构域与6个组氨酸（His）尾巴和谷胱甘肽-S-转移酶（GST）融合，进行纯化，然后利用His对Ni2+离子的亲和力，将其与镍-亚氨基二乙酸（Ni-NTA）柱结合。该柱用于分离对酵母SH3-Cdc25p结构域具有亲和力的酵母蛋白。如此分离出的主要蛋白经测序后被鉴定为酵母甘油醛-3-磷酸脱氢酶（GAP3DH）。
