Crystallization and preliminary X-ray analysis of the cAMP-dependent protein kinase catalytic subunit from Saccharomyces cerevisiae.
Crystallization and preliminary X-ray analysis of the cAMP-dependent protein kinase catalytic subunit from Saccharomyces cerevisiae.
复制标题
酿酒酵母 cAMP 依赖性蛋白激酶催化亚基的结晶和初步 X 射线分析。
DOI：
10.1021/bi00107a031
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发表时间：
1991
期刊：
影响因子：
2.9
通讯作者：
Pflugrath,JW
中科院分区：
文献类型：
--
作者：
Kuret,J;Pflugrath,JW
1991年8月12日收到的修订版摘要：TPK 1是酵母cAMP依赖性蛋白激酶催化亚基，其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下从2-丙醇和镁的溶液中结晶。晶体为六方双锥，空间群P6122（P6522），晶胞参数a= B= 61 A，c= 320 A。适用于衍射分析的大单晶可以通过微晶种获得，并且分辨率超过2.8-A。晶体密度测量显示每个单位细胞有12个激酶单体，每个不对称单位有一个激酶单体。1989年）。在真核细胞中大量的蛋白质磷酸化（Chelsky et al.，1985）指出了蛋白激酶超家族的存在，其每个成员都发挥重要的调节作用（Hunter，
Revised Manuscript Received August 12, 1991 abstract: 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 P6l22 (P6522), 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 densitymeasurements reveal 12 kinase monomers per unit cell with a single kinase monomer per asymmetric unit.I^ otein kinases are regulatory enzymes that catalyze the transfer of phosphate from ATP to protein substrates (Edelman et al., 1989). The large number of proteins phos-phorylated in eukaryotic cells (Chelsky et al., 1985) points to the existence of a protein kinase superfamily, each member of which carries out an important regulatory role (Hunter,