Topographical analysis of the plasma membrane-associated sucrose binding protein from soybean.
Plasma membranes of soybean cells actively engaged in sucrose transport have a sucrose binding protein (SBP) that does not appear to be an integral membrane protein. Experiments were undertaken to analyze the topographical association of this protein with the membrane. Treatment of purified plasma membrane vesicles with either 1 M KCl or KI released less than 35% of the sucrose binding protein from the membrane whereas treatment with either 4 M urea or 0.1 M Na2CO3, pH 11.5, disassociated between 50 and 70%, respectively, of this protein from the membrane. SDS, at either 0.5x, 1x, or 10x of its critical micelle concentration, effectively solubilized the sucrose binding protein. The nonionic detergents Triton X-100 and CHAPS, at either 0.5x, 1x, or 10x of their critical micelle concentration, solubilized between 65 and 75% of this protein. When either native plasma membrane-associated or in vitro-transcribed and -translated SBP were subjected to Triton X-114 phase separation, 80% partitioned into the detergent-poor aqueous phase. These results indicate that the SBP is a peripheral membrane protein but also suggest that there is a population of this protein that is tethered to the membrane.
Overvoorde
P J
PJ
Department of Botany, Washington State University, Pullman 99164-4238.
Grimes
H D
HD
D002352:Carrier Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D002462:Cell Membrane / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D026901:Membrane Transport Proteins; D037121:Plant Lectins; D010940:Plant Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D011487:Protein Conformation; D030262:Soybean Proteins; D013025:Glycine max / Q000737:chemistry*; D013395:Sucrose / Q000378:metabolism*
D002352:Carrier Proteins; D026901:Membrane Transport Proteins; D037121:Plant Lectins; D010940:Plant Proteins; D030262:Soybean Proteins; C078849:SBP protein, Glycine max; D013395:Sucrose