Membrane interactions influence the peptide binding behavior of DR1.
We analyzed the binding of an influenza matrix protein-derived peptide, MAT(17-31), to cell surface and purified DR1. The pH dependence of peptide binding was dramatically influenced by the membrane environment. Cell surface binding was enhanced at low pH, with little or no binding detected at neutral pH and optimal binding at pH 4. By contrast, hydrogen ion concentration had minimal effect on peptide binding to purified DR1. Exposure to low pH in the absence of peptide did not affect the peptide binding capacity of cell-associated DR1. Purified DR1 was stable at low pH, excluding the possibility that enhanced binding was offset by a competing denaturation event at low pH. The striking effect of pH on peptide binding characteristic of cell surface DR1 was recovered after reconstitution of purified DR1 in B cell membranes by detergent dialysis. This behavior was partially recovered by reconstitution of full-length, but not truncated DR1 in vesicles containing purified lipid. Our results demonstrate that interactions involving membrane components influence the peptide-binding behavior of DR1.
Sherman
M A
MA
Department of Pathology, Emory University School of Medicine, Atlanta, Georgia 30322.
Runnels
H A
HA
Moore
J C
JC
Stern
L J
LJ
Jensen
P E
PE
D000595:Amino Acid Sequence; D001665:Binding Sites; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000378:metabolism*; D010750:Phosphoproteins / Q000378:metabolism*; D011485:Protein Binding; D014157:Transcription Factors / Q000378:metabolism*; D014763:Viral Matrix Proteins / Q000378:metabolism*
D010446:Peptide Fragments; D010750:Phosphoproteins; D014157:Transcription Factors; D014763:Viral Matrix Proteins; C075995:down-regulator of transcription 1; C085077:influenza matrix peptide MAT(17-31)