Breaking the bottleneck: eukaryotic membrane protein expression for high-resolution structural studies.
The recombinant expression of eukaryotic membrane proteins has been a major stumbling block in efforts to determine their structures. In the last two years, however, five such proteins have yielded high-resolution X-ray or electron diffraction data, opening the prospect of increased throughput for eukaryotic membrane protein structure determination. Here, we summarize the major expression systems available, and highlight technical advances that should facilitate more systematic screening of expression conditions for this physiologically important class of targets.
Midgett
Charles R
CR
Department of Biochemistry, Dartmouth Medical School, 7200 Vail Building, Hanover, NH 03755, USA.
Madden
Dean R
DR
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D002460:Cell Line; D002846:Chromatography, Affinity; D003001:Cloning, Molecular / Q000379:methods*; D003461:Crystallography / Q000379:methods*; D018360:Crystallography, X-Ray; D004926:Escherichia coli; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D007313:Insecta / Q000166:cytology; D008322:Mammals; D008565:Membrane Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D046529:Microscopy, Electron, Transmission / Q000379:methods*; D018832:Molecular Chaperones / Q000502:physiology; D046912:Multiprotein Complexes; D011487:Protein Conformation; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D014162:Transfection / Q000379:methods; D015003:Yeasts
D008565:Membrane Proteins; D018832:Molecular Chaperones; D046912:Multiprotein Complexes; D011993:Recombinant Fusion Proteins