Department of Biochemistry, Mississippi State University, Mississippi State, Mississippi 39762.
Plant Physiol. 1987 Jun;84(2):337-40. doi: 10.1104/pp.84.2.337.
Oat (Avena sativa L.) seeds harvested at 2-day intervals from anthesis to maturity were tested for their ability to incorporate [(35)S]sulfate into protein. Incorporation of [(35)S]sulfate into TCA-insoluble material began 2 to 4 days postanthesis (DPA), reached a peak 14 to 16 DPA, and was barely detectable by 24 DPA. Incorporation of label into globulin was parallel to total protein accumulation, and averaged about 85% of the total protein synthesis. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of total protein extracted from developing seeds indicated that some polypeptides coinciding with the alpha and beta globulin subunits were present 2 to 4 DPA, but the full complement of globulin polypeptides was not present until 10 DPA. Immunoprecipitation of in vivo labeled seed extracts showed that globulin polypeptides and the 59 kilodalton precursor were present at early stages of development (4 DPA). Quantitation of dot blot analysis, using an oat globulin cDNA clone as a probe, indicated that one species of oat globulin mRNA was most abundant 15 DPA, which is during the peak time of storage protein synthesis.
从开花到成熟,每隔两天收获的燕麦(Avena sativa L.)种子被测试其将 [(35)S]硫酸盐掺入蛋白质的能力。[(35)S]硫酸盐掺入 TCA 不溶性物质始于开花后 2 至 4 天(DPA),在 14 至 16 DPA 达到峰值,在 24 DPA 时几乎无法检测到。标记物掺入球蛋白与总蛋白积累平行,平均约占总蛋白合成的 85%。从发育中的种子中提取的总蛋白的十二烷基硫酸钠-聚丙烯酰胺凝胶电泳表明,一些与 α 和 β 球蛋白亚基相吻合的多肽在 2 至 4 DPA 时存在,但直到 10 DPA 时才存在完整的球蛋白多肽。体内标记种子提取物的免疫沉淀显示,球蛋白多肽和 59 千道尔顿前体在发育的早期阶段(4 DPA)存在。使用燕麦球蛋白 cDNA 克隆作为探针的斑点印迹分析定量表明,一种燕麦球蛋白 mRNA 在 15 DPA 时最为丰富,此时是储存蛋白合成的高峰期。