Salmanowicz Bolesław P, Langner Monika, Kubicka-Matusiewicz Helena
Institute of Plant Genetics, Polish Academy of Sciences , PL 60-479 Poznań, Poland.
J Agric Food Chem. 2014 Oct 29;62(43):10535-41. doi: 10.1021/jf502926q. Epub 2014 Oct 20.
In this study, identification and characterization of the rye HMW secalin subunit (HMW-SS) composition in 68 inbred rye (Secale cereale L.) lines was performed by capillary zone electrophoresis (CZE). The HMW-SS were separated in an uncoated fused-silica capillary using an isoelectric iminodiacetic buffer in combination with poly(ethylene oxide), lauryl sulfobetaine, and acetonitrile as the separation buffer. The separations of the nonalkylated HMW-SS provided very good resolution and high reproducibility. Generally, the x-type rye HMW-SS were more abundant and have longer migration times than the y-type subunits. Both types of rye HMW-SS were separated into the major protein peak and one or two minor peaks. In total, seven x-type HMW-SS, five of which were newly identified subunits, and six y-type subunits, four of which were new, were distinguished on the basis of their CZE migration times. The migration order of the rye HMW-SS using CZE differed considerably from the relative electrophoretic mobilities in the SDS-PAGE gels.
在本研究中,采用毛细管区带电泳(CZE)对68个自交黑麦(Secale cereale L.)品系的黑麦高分子量麦醇溶蛋白亚基(HMW-SS)组成进行了鉴定和表征。使用未涂层熔融石英毛细管,以等电亚氨基二乙酸缓冲液与聚环氧乙烷、月桂基磺基甜菜碱和乙腈作为分离缓冲液,对HMW-SS进行分离。未烷基化HMW-SS的分离具有很好的分辨率和高重现性。一般来说,x型黑麦HMW-SS比y型亚基更丰富,迁移时间更长。两种类型的黑麦HMW-SS均被分离为主要蛋白峰和一个或两个次要峰。根据其CZE迁移时间,共区分出7个x型HMW-SS,其中5个是新鉴定的亚基,以及6个y型亚基,其中4个是新的。使用CZE时黑麦HMW-SS的迁移顺序与SDS-PAGE凝胶中的相对电泳迁移率有很大差异。