显微注射碱性磷酸酶及其低分子量底物对非洲爪蟾卵母细胞第一次减数分裂的体内效应。
In vivo effects of microinjected alkaline phosphatase and its low molecular weight substrates on the first meiotic cell division in Xenopus laevis oocytes.
作者信息
Hermann J, Mulner O, Bellé R, Marot J, Tso J, Ozon R
出版信息
Proc Natl Acad Sci U S A. 1984 Aug;81(16):5150-4. doi: 10.1073/pnas.81.16.5150.
Xenopus laevis oocytes were microinjected with low molecular weight phosphoesters such as 2-glycerophosphate, phosphotyrosine, phosphoserine, phosphothreonine, 4-nitrophenyl phosphate, and orthophosphate. These compounds were able to induce a considerable reduction in the time course of progesterone-induced maturation, with 2-glycerophosphate being the most effective. The basal level of cAMP and its drop during maturation were not affected by the microinjection of 2-glycerophosphate. The injection of alkaline phosphatase (EC 3.1.3.1.) from calf intestine at a low concentration (10 ng per oocyte) was able to decrease or abolish the effect of 2-glycerophosphate. At higher concentration (25 ng per oocyte) this enzyme totally blocked progesterone- or maturation-promoting factor-induced maturation. Alkaline phosphatase might behave in vivo as a phosphoprotein phosphatase active towards phosphotyrosine-containing proteins. In addition, our results indicate that phosphate or phosphoester-containing buffers should be avoided in the course of maturation-promoting factor purification.
将低分子量磷酸酯（如2-甘油磷酸酯、磷酸酪氨酸、磷酸丝氨酸、磷酸苏氨酸、4-硝基苯磷酸酯和正磷酸盐）显微注射到非洲爪蟾卵母细胞中。这些化合物能够显著缩短孕酮诱导成熟的时间进程，其中2-甘油磷酸酯最为有效。2-甘油磷酸酯的显微注射不影响cAMP的基础水平及其在成熟过程中的下降。以低浓度（每个卵母细胞10纳克）注射来自小牛肠的碱性磷酸酶（EC 3.1.3.1.）能够降低或消除2-甘油磷酸酯的作用。在较高浓度（每个卵母细胞25纳克）时，这种酶完全阻断孕酮或促成熟因子诱导的成熟。碱性磷酸酶在体内可能表现为对含磷酸酪氨酸的蛋白质有活性的磷蛋白磷酸酶。此外，我们的结果表明，在促成熟因子纯化过程中应避免使用含磷酸盐或磷酸酯的缓冲液。
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