Hutchison James M, Lu Zhenwei, Li Geoffrey C, Travis Benjamin, Mittal Ritesh, Deatherage Catherine L, Sanders Charles R
Department of Biochemistry, Vanderbilt University School of Medicine , Nashville, Tennessee 37240, United States.
Anatrace , 434 West Dussel Drive, Maumee, Ohio 43537, United States.
Biochemistry. 2017 Oct 17;56(41):5481-5484. doi: 10.1021/acs.biochem.7b00810. Epub 2017 Oct 9.
There remains a need for new non-ionic detergents that are suitable for use in biochemical and biophysical studies of membrane proteins. Here we explore the properties of n-dodecyl-β-melibioside (β-DDMB) micelles as a medium for membrane proteins. Melibiose is d-galactose-α(1→6)-d-glucose. Light scattering showed the β-DDMB micelle to be roughly 30 kDa smaller than micelles formed by the commonly used n-dodecyl-β-maltoside (β-DDM). β-DDMB stabilized diacylglycerol kinase (DAGK) against thermal inactivation. Moreover, activity assays conducted using aliquots of DAGK purified into β-DDMB yielded activities that were 40% higher than those of DAGK purified into β-DDM. β-DDMB yielded similar or better TROSY-HSQC NMR spectra for two single-pass membrane proteins and the tetraspan membrane protein peripheral myelin protein 22. β-DDMB appears be a useful addition to the toolbox of non-ionic detergents available for membrane protein research.
仍然需要适用于膜蛋白生化和生物物理研究的新型非离子洗涤剂。在这里,我们探索了正十二烷基-β-蜜二糖(β-DDMB)胶束作为膜蛋白介质的特性。蜜二糖是D-半乳糖-α(1→6)-D-葡萄糖。光散射显示β-DDMB胶束比常用的正十二烷基-β-麦芽糖苷(β-DDM)形成的胶束小约30 kDa。β-DDMB可稳定二酰基甘油激酶(DAGK)使其免于热失活。此外,使用纯化到β-DDMB中的DAGK等分试样进行的活性测定产生的活性比纯化到β-DDM中的DAGK高40%。对于两种单次跨膜蛋白和四跨膜蛋白外周髓磷脂蛋白22,β-DDMB产生了相似或更好的TROSY-HSQC NMR谱。β-DDMB似乎是可用于膜蛋白研究的非离子洗涤剂工具箱中的一个有用补充。