Hoyer L L, Cieslinski L B, McLaughlin M M, Torphy T J, Shatzman A R, Livi G P
Department of Gene Expression Sciences, SmithKline Beecham Pharmaceuticals, King of Prussia, Pennsylvania 19406.
Microbiology (Reading). 1994 Jul;140 ( Pt 7):1533-42. doi: 10.1099/13500872-140-7-1533.
We have cloned a Candida albicans gene, which encodes a cyclic nucleotide phosphodiesterase (PDEase), by complementation in a Saccharomyces cerevisiae PDEase-deficient mutant. The deduced amino acid sequence is similar to that of the low-affinity PDEase of S. cerevisiae (PDE1) and the cyclic nucleotide PDEase (PD) of Dictyostelium discoideum. Biochemical analysis of recombinant protein produced in S. cerevisiae indicated that the enzyme behaves as a PDE1 homologue: it hydrolyses both cAMP (Km = 0.49 mM) and cGMP (Km = 0.25 mM), does not require divalent cations for maximal activity and is only moderately inhibited by millimolar concentrations of standard PDEase inhibitors. Based on these data, we designate the C. albicans we have cloned, PDE1. Low-stringency genomic Southern blots showed cross-hybridization between C. albicans PDE1 and DNA from Candida stellatoidea, but not with DNA from S. cerevisiae or several closely related Candida species.
我们通过在酿酒酵母磷酸二酯酶缺陷型突变体中进行互补克隆了一个白色念珠菌基因，该基因编码一种环核苷酸磷酸二酯酶（PDEase）。推导的氨基酸序列与酿酒酵母的低亲和力磷酸二酯酶（PDE1）和盘基网柄菌的环核苷酸磷酸二酯酶（PD）相似。对酿酒酵母中产生的重组蛋白进行生化分析表明，该酶表现为PDE1同源物：它水解cAMP（Km = 0.49 mM）和cGMP（Km = 0.25 mM），最大活性不需要二价阳离子，并且仅受到毫摩尔浓度的标准磷酸二酯酶抑制剂的中度抑制。基于这些数据，我们将我们克隆的白色念珠菌命名为PDE1。低严谨度基因组Southern印迹显示白色念珠菌PDE1与星状念珠菌的DNA之间存在交叉杂交，但与酿酒酵母或几种密切相关的念珠菌属的DNA没有交叉杂交。
Zotero 插件