NMR Characterization of Recombinant Transmembrane Protein CB2 Fragment CB2 180-233

Title: NMR Characterization of Recombinant Transmembrane Protein CB2 Fragment CB2 180-233

Volume: 13 Issue: 4

Author(s): J. Zhao, H. Zheng and X.- Q. Xie

Affiliation:

Keywords: GPCR, CB2, transmembrane helical domain, NMR, membrane protein

Abstract: The expression of membrane proteins has been the bottleneck for their structural studies. Recently, we developed a method to obtain milligram quantities of isotope-labeled seven transmembrane G-protein coupled cannabinoid (CB) receptor fragment in E. coli. In order to verify this method and confirm the recombinant isotope-labeled CB2 fragment, 3D hetero-nuclear NMR techniques were used to analyze the structure of the fragment CB2 180-233 in DMSO-d6 solvent. The sequential assignments of TM5 and intra-cellular loop 3 were accomplished, which confirmed the experimental protocols of isotope-labeled recombinant protein expression, fusion protein cleavage, and membrane protein purification. The obtained structure also showed α-helix in the TM5 region, but it was interrupted by a disordered region (Gly204_ILe206). These results further revealed that our established approach is a promising method to express recombinant membrane proteins for their structural studies.

Cite this article as:

Zhao J., Zheng H. and Xie Q. X.-, NMR Characterization of Recombinant Transmembrane Protein CB2 Fragment CB2 180-233, Protein & Peptide Letters 2006; 13 (4) . https://dx.doi.org/10.2174/092986606775974483