Kuret J, Pflugrath J W
Cold Spring Harbor Laboratory, Cold Spring Harbor, New York 11724.
Biochemistry. 1991 Oct 29;30(43):10595-600. doi: 10.1021/bi00107a031.
A truncated variant of TPK1, the yeast cAMP-dependent protein kinase catalytic subunit, was overexpressed in an engineered strain of Saccharomyces cerevisiae, purified by liquid chromatography, and crystallized from solutions of 2-propanol and magnesium at alkaline pH. The crystals are hexagonal dipyramids, space group P6(1)22 (P6(5)22), with unit-cell parameters a = b = 61 A, c = 320 A. Large single crystals suitable for diffraction analysis are obtainable by microseeding, and diffract beyond 2.8-A resolution. Crystal density measurements reveal 12 kinase monomers per unit cell with a single kinase monomer per asymmetric unit.
酵母环磷酸腺苷依赖性蛋白激酶催化亚基TPK1的截短变体在酿酒酵母工程菌株中过表达，通过液相色谱法纯化，并在碱性pH条件下从异丙醇和镁的溶液中结晶。晶体为六方双锥体，空间群P6(1)22（P6(5)22），晶胞参数a = b = 61 Å，c = 320 Å。通过微量接种可获得适合衍射分析的大单晶，其衍射分辨率超过2.8 Å。晶体密度测量显示每个晶胞中有十二个激酶单体，每个不对称单元中有一个激酶单体。
