Quantitative assay of cyclin-dependent kinase activity as a sensitive marker of cell proliferation in marine teleost larvae

被引:5
作者
Westerman, ME
Holt, GJ
DiMichele, L
机构
[1] Texas A&M Univ, Dept Wildlife & Fisheries, College Stn, TX 77843 USA
[2] Univ Texas, Inst Marine Sci, Port Aransas, TX 78373 USA
关键词
cell proliferation; Cdks; MPF; suc1; growth; teleost;
D O I
10.1007/PL00011779
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
An evolutionarily conserved family of cyclin-dependent kinases (Cdks) regulates cell cycle progression in eukaryotes. Mitosis-promoting factor (MPF) is a transiently activated Cdk required for mitosis. We propose use of partially purified MPF kinase activity as a sensitive marker of cell proliferation in marine teleost larvae and describe a quantitative spin-filter assay of suc1-precipitated MPF activity. MPF extracted from embryos and larvae of red drum, Sciaenops ocellatus, phosphorylated histone H1 protein and a Cdk-specific peptide substrate. Kinetic analyses demonstrated that apparent K-m values of red drum MPF for adenosine triphosphate and the peptide substrate were 98.3 and 18.2 mu M, respectively. A simple growth experiment showed that MPF activity was significantly greater in rapidly growing red drum larvae than in intermittently fed cohorts growing more slowly.
引用
收藏
页码:297 / 310
页数:14
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