Quantitation of LMW-GS to HMW-GS Ratio in Wheat Flours
Quantitation of LMW-GS to HMW-GS Ratio in Wheat Flours
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DOI：
10.1094/cchem-85-6-0824
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发表时间：
2008-11-01
期刊：
影响因子：
2.4
通讯作者：
MacRitchie, Finlay
中科院分区：
文献类型：
--
作者：
Cinco-Moroyoqui, Francisco J.;MacRitchie, Finlay
对谷蛋白低分子量/高分子量亚基（GS）比值的测定方法进行了比较。利用一组HMW-GS数目在0 - 5之间的近等基因系，提供了宽范围的LMW/HMW-GS。麦谷蛋白的制备是在用二甲基亚砜（DMSO）或50%丙醇溶解单体蛋白质后，或通过在制备型SE-HPLC柱前收集馏分，从圆形全麦粉中获得的。通过RP-HPLC、SE-HPLC和SDS-PAGE对来自三种制备物中的每一种的还原谷蛋白进行分析。两种溶剂，DMSO和50%丙醇。提取了大量的聚合蛋白质，因此对测定的准确性产生了一些怀疑。当在制备型SE-HPLC柱上收集总谷蛋白提取物的洗脱液之前收集聚合级分时，该问题在很大程度上得以避免。两种溶剂对具有较多HMW-GS或具有ID染色体编码的强度相关HMW-GS 5+10的品系去除较少的谷蛋白。采用SE-HPLC收集多聚体蛋白，再用RP-HPLC分离谷蛋白亚基，是测定LMW/HMW-GS比值的最佳方法。SE-HPLC分离了两组亚基以及HMW白蛋白。RP-HPLC具有能够定量单个亚基的潜在优势。
A comparison was made of methods for measuring the LMW/HMW glutenin subunit (GS) ratio for glutenin. A set of near-isogenic wheat lines With the number of HMW-GS varying from 0 to 5 was utilized to provide a wide range of LMW/HMW-GS. Glutenin preparations were obtained from,round whole meal after solubilization of monomeric proteins by dimethyl sulfoxide (DMSO) or 50% propanol or by fraction collection front a preparative SE-HPLC column. Analyses were made oil the reduced glutenin from each of the three preparations by RP-HPLC, SE-HPLC, and SDS-PAGE. Both solvents, DMSO and 50% propanol. extracted appreciable amounts of polymeric protein, thus casting some doubts on the accuracy of the determinations. This problem was largely avoided when the polymeric fraction was collected front the eluate of a total glutenin extract run oil a preparative SE-HPLC column. Less glutenin was removed by the two solvents for lines with a greater number of HMW-GS or with strength-associated HMW-GS 5+10 coded by the ID chromosome. Collection of the polymeric protein in SE-HPLC, followed by separation of the glutenin subunits in RP-HPLC, was the best method for quantitating the LMW/HMW-GS ratio. SE-HPLC gave a separation of the two groups of subunits as well as HMW albumins. RP-HPLC has the potential advantage of being able to quantitate individual subunits.