The role of the N-terminal domain of the complement fragment receptor C5L2 in ligand binding.
C5L2 is a new cellular receptor found to interact with the human anaphylatoxins complement factor C5a and its C-terminal cleavage product C5a des Arg. The classical human C5a receptor (C5aR) preferentially binds C5a, with a 10-100-fold lower affinity for C5a des Arg. In contrast, C5L2 binds both ligands with nearly equal affinity. C5aR presents acidic and tyrosine residues in its N terminus that interact with the core of C5a while a hydrophobic pocket formed by the transmembrane helices interacts with residues in the C terminus of C5a. Here, we have investigated the molecular basis for the increased affinity of C5L2 for C5a des Arg. Rat and mouse C5L2 preferentially bound C5a des Arg, whereas rodent C5aR showed much higher affinity for intact C5a. Effective peptidic and non-peptidic ligands for the transmembrane hydrophobic pocket of C5aR were poor inhibitors of ligand binding to C5L2. An antibody raised against the N terminus of human C5L2 did not affect the binding of C5a to C5L2 but did inhibit C5a des Arg binding. A chimeric C5L2, containing the N terminus of C5aR, had little effect on the affinity for C5a des Arg. Mutation of acidic and tyrosine residues in the N terminus of human C5L2 revealed that 3 residues were critical for C5a des Arg binding but had little involvement in C5a binding. C5L2 thus appears to bind C5a and C5a des Arg by different mechanisms, and, unlike C5aR, C5L2 uses critical residues in its N-terminal domain for binding only to C5a des Arg.
Scola
Anne-Marie
AM
Academic Neurology Unit and Department of Molecular Biology and Biotechnology, University of Sheffield, Sheffield S10 2RX, United Kingdom.
Higginbottom
Adrian
A
Partridge
Lynda J
LJ
Reid
Robert C
RC
Woodruff
Trent
T
Taylor
Stephen M
SM
Fairlie
David P
DP
Monk
Peter N
PN
D019943:Amino Acid Substitution / Q000235:genetics; D000818:Animals; D001665:Binding Sites / Q000235:genetics; D016466:CHO Cells; D045744:Cell Line, Tumor; D015937:Complement C5a, des-Arginine / Q000378:metabolism; D006224:Cricetinae; D003412:Cricetulus; D006801:Humans; D008024:Ligands; D051379:Mice; D010446:Peptide Fragments / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D011485:Protein Binding / Q000235:genetics; D051381:Rats; D044087:Receptor, Anaphylatoxin C5a / Q000235:genetics / Q000378:metabolism / Q000502:physiology*
C496868:C5ar2 protein, rat; D015937:Complement C5a, des-Arginine; D008024:Ligands; D010446:Peptide Fragments; D044087:Receptor, Anaphylatoxin C5a