Similar binding properties of peptide ligands for a human immunoglobulin and its light chain dimer.
The urinary light chain dimer and serum monoclonal IgG1 protein from a patient (Mcg) with multiple myeloma and amyloidosis were systematically tested for their binding activities to peptides presented on solid supports. The system was validated using a series of enkephalins, beta-casomorphins and DNP-lysine derivatives which were known to complex with the dimer. Sets of peptide ligands binding to the proteins were constructed by incremental additions of amino acid residues to minimal binding units [Geysen et al., J. Immun. Meth. 102, 259-274 (1987)]. Both the amino acid sequences and the combinations of optical isomers were optimized at each stage of the syntheses. Binding could be demonstrated for ligands ranging in size from a tethered single amino acid to pentapeptides. At the dipeptide levels, the dimer and the IgG1 protein showed different preferences (Hp versus qf, where lower case letters designate D-amino acid residues). However, in a tetrapeptide ligand (qfHp) for the dimer, both of these initial preferences had converged. With few exceptions, the IgG1 molecule showed binding activity for the ligands developed for the dimer. Two sets of selected peptides, one based on Hp and the other on mW, were synthesized for diffusion into crystals of the dimer. X-ray analyses showed that these peptides bound exclusively in the main binding cavity between the "variable" domains of the dimer. As predicted from the ELISA results with tethered ligands, the relative occupancies in the crystals followed the order of tetrapeptide greater than tripeptide much greater than dipeptide. The crystallographic studies confirmed that peptides with very different sequences can bind in the same cavity.
Tribbick
G
G
Coselco Mimotopes Pty Ltd, Commonwealth Serum Laboratories, Parkville, Victoria, Australia.
Edmundson
A B
AB
Mason
T J
TJ
Geysen
H M
HM
D001536:Bence Jones Protein; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D004136:Dinitrobenzenes; D004723:Endorphins; D004745:Enkephalins; D005973:Glutamine; D006639:Histidine; D006801:Humans; D007074:Immunoglobulin G; D008024:Ligands; D009194:Myeloma Proteins; D010455:Peptides
D004136:Dinitrobenzenes; D004723:Endorphins; D004745:Enkephalins; D007074:Immunoglobulin G; D008024:Ligands; D009194:Myeloma Proteins; D010455:Peptides; D005973:Glutamine; D006639:Histidine; C021968:beta-casomorphins; D001536:Bence Jones Protein