Detergent Screening for NMR-Based Structural Study of the Integral Membrane Protein, Emopamil Binding Protein (Human Sterol Δ8-Δ7 Isomerase)
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초록
Human sterol Δ8-Δ7 isomerase, commonly known as emopamil binding protein (EBP), is an essential protein in the cholesterol-synthetic pathway, and mutations of this protein are critically associated with human diseases such as Conradi-Hunermann–Happle or male EBP disorder with neurological defects syndrome. Due to such a clinical importance, EBP has been intensively investigated and some important features have been reported. EBP is a tetra-spanning membrane protein, of which 2nd, 3rd, and 4th membrane-spanning α helices play an important role in its enzymatic function. However, detailed structural feature at atomic resolution has not yet been elucidated, due to characteristic difficulties in dealing with membrane protein. Here, we over-expressed EBP using Escherichia coli and performed detergent screening to find suitable membrane mimetics for structural studies of the protein by NMR. As results, DPC and LMPG could be evaluated as the most favorable detergents to acquire promising NMR spectra for structural study of EBP.
Abstract
Human sterol Δ8-Δ7 isomerase, commonly known as emopamil binding protein (EBP), is an essential protein in the cholesterol-synthetic pathway, and mutations of this protein are critically associated with human diseases such as Conradi-Hunermann–Happle or male EBP disorder with neurological defects syndrome. Due to such a clinical importance, EBP has been intensively investigated and some important features have been reported. EBP is a tetra-spanning membrane protein, of which 2nd, 3rd, and 4th membrane-spanning α helices play an important role in its enzymatic function. However, detailed structural feature at atomic resolution has not yet been elucidated, due to characteristic difficulties in dealing with membrane protein. Here, we over-expressed EBP using Escherichia coli and performed detergent screening to find suitable membrane mimetics for structural studies of the protein by NMR. As results, DPC and LMPG could be evaluated as the most favorable detergents to acquire promising NMR spectra for structural study of EBP.
참고문헌 (27)
- L. X. Finegold, "Cholesterol in membrane models" (1992), CRC Press
- P. L. Yeagle, , Biochimie, 73, pp.1303~ (1991)
- P. L. Yeagle, , Biochim. Biophys. Acta, 822, pp.267~ (1985)
- L. J. Sharpe, , J. Biol. Chem, 288, pp.18707~ (2013)
- A. A. Kandutsch, , J. Biol. Chem, 235, pp.2256~ (1960)
- K. Bloch, , Steroids, 57, pp.378~ (1992)
- L. V. Furtado, , Am. J. Med. Genet, 152A, pp.2838~ (2010)
- A. Kolb-Maurer, , Acta Derm-Venereol, 88, pp.47~ (2008)
- B. U. Fitzky, , Proc. Natl. Acad. Sci. U. S. A, 95, pp.8181~ (1998)
- F. F. Moebius, , Mol. Pharmacol, 43, pp.139~ (1993)
- F. F. Moebius, , J. Biol. Chem, 269, pp.29314~ (1994)
- M. Hanner, , J. Biol. Chem, 270, pp.7551~ (1995)
- F. F. Moebius, , Mol. Pharmacol, 54, pp.591~ (1998)
- S. Silve, , J. Biol. Chem, 271, pp.22434~ (1996)
- S. Silve, , Mol. Cell. Biol, 16, pp.2719~ (1996)
- F. F. Moebius, , Biochemistry, 38, pp.1119~ (1999)
- A. Rahier, , Biochem. J, 414, pp.247~ (2008)
- A. Korepanova, , Protein Sci, 14, pp.148~ (2005)
- M. Renault, , Methods Mol. Biol, 654, pp.321~ (2010)
- B. Liang, , Proc. Natl. Acad. Sci. U. S. A, 104, pp.16140~ (2007)
- R. C. Page, , J. Struct. Funct. Genomics, 7, pp.51~ (2006)
- 심대원, "A simple guide to the structural study on membrane proteins in detergents using solution NMR", Journal of the Korean Magnetic Resonance Society, 19(3), pp.137~142 (2015), 한국자기공명학회[DOI]
- 백수연, "Improved Purification of Thermophilic FoF1-ATPSynthase c-Subunit Rings and Solid-State NMR Characterization of Them in Different Lipid Membranes", Journal of the Korean Magnetic Resonance Society, 17(2), pp.67~75 (2013), 한국자기공명학회
- L. Columbus, , J. Am. Chem. Soc, 131, pp.7320~ (2009)
- D. Nietlispach, , J. Biomol. NMR, 31, pp.161~ (2005)
- F. Delaglio, , J. Biomol. NMR, 6, pp.277~ (1995)
- T. Goddard, "SPARKY 3" (2008), University of California