Amir Mohd, Dar Mohammad Aasif, Islam Asimul, Ahmad Faizan, Imtaiyaz Hassan Md
Centre for Interdisciplinary Research in Basic Sciences, Jamia Millia Islamia, New Delhi, 110025, India.
3 Biotech. 2016 Jun;6(1):85. doi: 10.1007/s13205-016-0400-3. Epub 2016 Mar 1.
Rho GTPases activating protein 2 (RGA2) is primarily involved in the modulation of numerous morphological events in eukaryotes. It protects plants by triggering the defense system which restricts the pathogen growth. This is the first report on the isolation, purification and characterization of RGA2 from the stems of Tinospora cordifolia, a medicinal plant. The RGA2 was purified using simple two-step process using DEAE-Hi-Trap FF and Superdex 200 chromatography columns, with a high yield. The purity of RGA2 was confirmed by SDS-PAGE and identified by MALDI-TOF/MS. The purified protein was further characterized for its secondary structural elements using the far-UV circular dichroism measurements. Our purification procedure is simple two-step process with high yield which can be further used to produce RGA2 for structural and functional studies.
Rho GTP酶激活蛋白2（RGA2）主要参与真核生物中众多形态学事件的调控。它通过触发限制病原体生长的防御系统来保护植物。这是关于从药用植物心叶青牛胆茎中分离、纯化和鉴定RGA2的首次报道。使用DEAE-Hi-Trap FF和Superdex 200色谱柱，通过简单的两步法纯化RGA2，产率很高。通过SDS-PAGE确认RGA2的纯度，并通过MALDI-TOF/MS进行鉴定。使用远紫外圆二色性测量对纯化后的蛋白质的二级结构元件进行进一步表征。我们的纯化程序是简单的两步法，产率高，可进一步用于生产RGA2以进行结构和功能研究。
