Detergent-enhanced dissociation of endogenous peptides from PI-DRB1*0401.
A variety of detergents have been shown to catalyze the dissociation of bound peptides from a soluble from of DRB1*0401. By using a class II molecule lacking the hydrophobic transmembrane region, the need for solubilizing the transmembrane protein was removed and enabled the specific interaction between the class II protein and the amphiphile to be identified. The presence of detergent increased the rate of association of added peptide and the percent occupancy of the receptor, presumably because the dissociation of endogenous peptide was the rate-limiting step in binding. The data help explain the differences reported between peptide binding to class II proteins on the surface of cells and binding to class II proteins solubilized in detergent. The interaction did not correlate with the critical micellar concentration of the detergent nor were all amphiphilic structures equally effective, consistent with a specific interaction between the amphiphile and the MHC class II protein. Of the eight detergents examined, octyl glucoside was the most efficient. These experiments did not distinguish between an allosteric mechanism or direct competition with the peptide for binding.
Buelow
R
R
ImmuLogic Pharm Corp., Palo Alto, CA 94304.
Kuo
S
S
Paborsky
L
L
Wilson
K J
KJ
Rothbard
J B
JB
D000595:Amino Acid Sequence; D003902:Detergents / Q000737:chemistry*; D005960:Glucosides / Q000737:chemistry; D006684:HLA-DR Antigens / Q000737:chemistry* / Q000378:metabolism; D059811:HLA-DRB1 Chains; D000949:Histocompatibility Antigens Class II / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D008823:Micelles; D008969:Molecular Sequence Data; D010455:Peptides / Q000276:immunology*; D010716:Phosphatidylinositols / Q000378:metabolism
D003902:Detergents; D005960:Glucosides; D006684:HLA-DR Antigens; D059811:HLA-DRB1 Chains; C553505:HLA-DRB1*04:01 antigen; D000949:Histocompatibility Antigens Class II; D008823:Micelles; D010455:Peptides; D010716:Phosphatidylinositols; C018619:octyl-beta-D-glucoside