燕麦根膜等电聚焦。
Isoelectric focusing of oat root membranes.
机构信息
Biology Department, Williams College, Williamstown, Massachusetts 01267.
出版信息
Plant Physiol. 1986 Sep;82(1):327-9. doi: 10.1104/pp.82.1.327.
The feasibility of purifying subcellular membranes, especially plasma membranes, from oat roots using isoelectric focusing has been examined. Membranes from oat (Avena sativa L. cv Garry) root homogenates were fractionated using discontinuous sucrose density gradient centrifugation and then electrofocused using a microanalytical isoelectric focusing column. The column contained either a broad-range (pH 3-10) or narrow-range (pH 3-6) pH gradient stabilized by a 5 to 15% Ficoll gradient. Results from the broad-range columns confirmed that the isoelectric pH (pI) values of the membranes were in the acidic range, with pI values ranging from 3.9 to 5.2. Using narrow-range pH gradients, it was possible to fractionate further plasma membrane-enriched material obtained from a sucrose density gradient. We had no success at fractionating crude membrane preparations from oat roots. Narrow-range pH gradients generated by commercial ampholytes were more successful than those generated by acetate/acetic acid mixtures.
已经研究了使用等电聚焦从燕麦根中纯化亚细胞膜（特别是质膜）的可行性。使用不连续蔗糖密度梯度离心从燕麦（Avena sativa L. cv Garry）根匀浆中分离膜，然后使用微分析等电聚焦柱进行电聚焦。该柱含有由 5%至 15%Ficoll 梯度稳定的宽范围（pH 3-10）或窄范围（pH 3-6）pH 梯度。宽范围柱的结果证实，膜的等电 pH（pI）值处于酸性范围，pI 值范围为 3.9 至 5.2。使用窄范围 pH 梯度，可以进一步分离从蔗糖密度梯度获得的富含质膜的物质。我们未能从燕麦根中分离粗膜制剂。由商业两性电解质产生的窄范围 pH 梯度比由乙酸盐/乙酸混合物产生的 pH 梯度更成功。
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