Reconstitution of full-length human caveolin-1 into phospholipid bicelles: Validation by analytical ultracentrifugation.
Reconstitution of full-length human caveolin-1 into phospholipid bicelles: Validation by analytical ultracentrifugation.
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DOI：
10.1016/j.bpc.2020.106339
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发表时间：
2020-04
影响因子：
3.8
通讯作者：
Glover KJ
中科院分区：
文献类型：
--
作者：
Rieth MD;Root KT;Glover KJ
A significant hurdle in obtaining biophysical information on membrane proteins is developing a successful strategy for their reconstitution into a suitable membrane mimic. In particular, utilization of the more ‘native-like’ membrane mimics such as bicelles is generally more challenging than simple micellar solubilization. Caveolin-1, an integral membrane protein involved in membrane curvature, endocytosis, mechano-protection, and signal transduction, has been shown to be particularly recalcitrant to standard reconstitution protocols due to its highly hydrophobic characteristics. Herein we describe a robust method to incorporate recombinantly produced full-length caveolin-1 into bicelles at levels needed for biophysical experimentation. The benchmark of successful reconstitution is the obtainment of protein in a homogeneous state; therefore, we developed a validation procedure to monitor the success of the reconstitution using analytical ultracentrifugation of density-matched bicelles. Our findings indicated that our protocol produces a very homogeneous preparation of caveolin-1 associated with bicelles, and that caveolin-1 is highly α-helical (by circular dichroism spectroscopy). We believe that this methodology will serve as a general strategy to facilitate biophysical studies on membrane proteins.
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影响因子：
2.9
作者：
Hoop, Cody L.;Sivanandam, V. N.;Kodali, Ravindra;Srnec, Matthew N.;van der Wel, Patrick C. A.
通讯作者：
van der Wel, Patrick C. A.
DOI：
10.1021/acs.jpclett.8b02079
发表时间：
2018-08-02
期刊：
The journal of physical chemistry letters
影响因子：
--
作者：
Caldwell TA;Baoukina S;Brock AT;Oliver RC;Root KT;Krueger JK;Glover KJ;Tieleman DP;Columbus L
通讯作者：
Columbus L
影响因子：
5.7
作者：
Laguerre, Aisha;Loehr, Frank;Doetsch, Volker
通讯作者：
Doetsch, Volker
影响因子：
2.9
作者：
Pike, LJ;Han, XL;Gross, RW
通讯作者：
Gross, RW
DOI：
10.1073/pnas.172196599
发表时间：
2002-08-20
影响因子：
11.1
作者：
Fernandez, I;Ying, YS;Anderson, RGW
通讯作者：
Anderson, RGW