Natural catalytic antibodies: peptide-hydrolyzing activities of Bence Jones proteins and VL fragment.
Monoclonal human light chains, i.e. Bence Jones proteins, and their recombinant variable fragments (VL) were screened for proteolytic activity using peptide-methylcoumarinamide (peptide-MCA) conjugates and vasoactive intestinal polypeptide (VIP) as substrates. Sixteen of 21 Bence Jones proteins and one of three VL fragments were capable of detectable cleavage of one or more substrates. The magnitude and kinetic characteristics of the activity varied with different substrates. Among the peptide-MCA substrates, the presence of tripeptide or tetrapeptide moieties with a basic residue at the scissile bond generally favored expression of the activity. The influence of N-terminal flanking residue recognition was evident from differing values of Km and kcat (turnover number) observed using different Arg-containing peptide-MCA substrates. Different light chains displayed different kinetic parameters for the same substrate, suggesting unique catalytic sites. Hydrolysis of VIP was characterized by nanomolar Michaelis-Menten constants (Km), suggesting comparatively high affinity recognition of this peptide. The 25-kDa monomer and the 50-kDa dimer forms of one light chain preparation were resolved by gel filtration in 6 M guanidine hydrochloride. Following renaturation, the monomer displayed 51-fold greater peptide-MCA-hydrolyzing activity than the dimer. A renatured VL domain prepared by gel filtration in 6 M guanidine hydrochloride displayed VIP-hydrolyzing activity in the 12.5-kDa peak fractions. These results provide evidence for the proteolytic activity of certain human light chains and imply that this phenomenon may have a pathophysiological significance.
Paul
S
S
Department of Anesthesiology, University of Nebraska Medical Center, Omaha 68198-6830, USA.
Li
L
L
Kalaga
R
R
Wilkins-Stevens
P
P
Stevens
F J
FJ
Solomon
A
A
D000595:Amino Acid Sequence; D001536:Bence Jones Protein / Q000378:metabolism*; D002384:Catalysis; D003374:Coumarins; D010450:Endopeptidases / Q000378:metabolism*; D006801:Humans; D007145:Immunoglobulin kappa-Chains / Q000378:metabolism*; D007146:Immunoglobulin lambda-Chains / Q000378:metabolism*; D007700:Kinetics; D008969:Molecular Sequence Data; D009101:Multiple Myeloma / Q000276:immunology / Q000652:urine; D009842:Oligopeptides / Q000378:metabolism; D010446:Peptide Fragments / Q000378:metabolism; D013379:Substrate Specificity; D014660:Vasoactive Intestinal Peptide / Q000378:metabolism
D003374:Coumarins; D007145:Immunoglobulin kappa-Chains; D007146:Immunoglobulin lambda-Chains; D009842:Oligopeptides; D010446:Peptide Fragments; D014660:Vasoactive Intestinal Peptide; D001536:Bence Jones Protein; D010450:Endopeptidases