The influence of detergents and amphiphiles on the solubility of the light-harvesting I complex.
The effect of detergents and amphiphiles on protein solubility and their use in crystallization solutions was examined for an integral membrane protein, the light-harvesting I complex from Rhodospirillum centenum. Measurement by a centrifugation assay of the solubility of the protein in different detergents and amphiphiles showed high protein-solubility values when either octyl glucoside or lauryldimethylamine-N-oxide was present with heptanetriol or when deoxycholate was present with spermine. The detergent/amphiphile combinations that resulted in high protein solubility were shown to be successful for crystallization of the protein, suggesting that crystallization is favored for detergents and amphiphiles that optimize the solubility of integral membrane proteins. The amphiphiles effective for crystallization were found using laser mass spectrometry to displace the lauryldimethylamine-N-oxide bound to the protein. These results suggest that mass spectrometry can be used for screening of favorable crystallization conditions.
Rosenow
M A
MA
Department of Chemistry and Biochemistry, Arizona State University, Tempe, AZ 85287-1604, USA.
Brune
D
D
Allen
J P
JP
D002498:Centrifugation; D003460:Crystallization; D003840:Deoxycholic Acid / Q000737:chemistry; D003902:Detergents / Q000494:pharmacology*; D004123:Dimethylamines / Q000737:chemistry; D005233:Fatty Alcohols / Q000737:chemistry; D005960:Glucosides / Q000737:chemistry; D007834:Lasers; D045342:Light-Harvesting Protein Complexes; D013058:Mass Spectrometry; D011506:Proteins / Q000737:chemistry; D042182:Rhodospirillum centenum / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013096:Spermine / Q000737:chemistry
D003902:Detergents; D004123:Dimethylamines; D005233:Fatty Alcohols; D005960:Glucosides; D045342:Light-Harvesting Protein Complexes; D011506:Proteins; D003840:Deoxycholic Acid; C068115:heptane-1,2,3-triol; C018619:octyl-beta-D-glucoside; D013096:Spermine; C014518:dodecyldimethylamine oxide