Enhanced solubilization of membrane proteins by alkylamines and polyamines.
Around 25% of proteins in living organisms are membrane proteins that perform many critical functions such as synthesis of biomolecules and signal transduction. Membrane proteins are extracted from the lipid bilayer and solubilized with a detergent for biochemical characterization; however, their solubilization is an empirical technique and sometimes insufficient quantities of proteins are solubilized in aqueous buffer to allow characterization. We found that addition of alkylamines and polyamines to solubilization buffer containing a detergent enhanced solubilization of membrane proteins from microsomes. The solubilization of polygalacturonic acid synthase localized at the plant Golgi membrane was enhanced by up to 9.9-fold upon addition of spermidine to the solubilization buffer. These additives also enhanced the solubilization of other plant membrane proteins localized in other organelles such as the endoplasmic reticulum and plasma membrane as well as that of an animal Golgi-localized membrane protein. Thus, addition of alkylamines and polyamines to solubilization buffer is a generally applicable method for effective solubilization of membrane proteins. The mechanism of the enhancement of solubilization is discussed.
Yasui
Kazutoshi
K
Department of Chemistry, Graduate School of Science, Osaka University, Toyonaka, Osaka 560-0043, Japan.
Uegaki
Masamichi
M
Shiraki
Kentaro
K
Ishimizu
Takeshi
T
D000588:Amines / Q000737:chemistry*; D000818:Animals; D002417:Cattle; D002462:Cell Membrane / Q000201:enzymology*; D003576:Electron Transport Complex IV / Q000737:chemistry; D008025:Ligases / Q000737:chemistry; D008565:Membrane Proteins / Q000737:chemistry*; D009245:NADH Dehydrogenase / Q000737:chemistry; D010368:Pectins / Q000096:biosynthesis; D010940:Plant Proteins / Q000737:chemistry; D011073:Polyamines / Q000737:chemistry*; D012995:Solubility; D005723:gamma-Glutamyltransferase / Q000737:chemistry
D000588:Amines; D008565:Membrane Proteins; D010940:Plant Proteins; D011073:Polyamines; D010368:Pectins; D009245:NADH Dehydrogenase; D003576:Electron Transport Complex IV; D005723:gamma-Glutamyltransferase; D008025:Ligases; C003181:polygalacturonic acid