Differences in the biochemical properties of esterolytic antibodies correlate with structural diversity.
A prerequisite to the design and engineering of catalytic antibodies is the knowledge of their structure and in particular which residues are involved in binding and catalysis. We compared the structure and catalytic properties of a series of six monoclonal antibodies which were all raised against a p-nitrophenyl (PNP) phosphonate and which catalyze the hydrolysis of p-nitrophenyl esters. Three of the antibodies (Group I) have similar light and heavy chain variable regions. The other three antibodies have similar VL regions of which two (Group II) have VH regions from the MOPC21 gene family and the remaining one (Group III) a VH from the MC101 gene family making a total of three different groups based on their V region sequences. The structural division into groups is paralleled by the differences in binding constants to hapten analogs, substrate specificity and the susceptibility of the catalytic activity of the antibodies to chemical modification of tryptophan and arginine residues. The relative binding of a transition state analog to the binding of substrate is much higher for the Group I antibodies than for the other groups. Only the Group I antibodies can catalyze the hydrolysis of a carbonate substrate. However all of the antibodies lose catalytic activity upon specific tyrosine modification which highlights the importance of tyrosine in the active site of the antibodies. Thus, antibodies raised against a single hapten can give antibodies with different structures, and correspondingly different specificities and catalytic properties.
Zemel
R
R
Department of Chemical Immunology, Weizmann Institute of Science, Rehovot, Israel.
Schindler
D G
DG
Tawfik
D S
DS
Eshhar
Z
Z
Green
B S
BS
D000595:Amino Acid Sequence; D000818:Animals; D017151:Antibodies, Catalytic / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA; D004952:Esters / Q000378:metabolism; D007143:Immunoglobulin Heavy Chains / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D007147:Immunoglobulin Light Chains / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D007135:Immunoglobulin Variable Region / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D007700:Kinetics; D051379:Mice; D008969:Molecular Sequence Data; D009596:Nitrophenols; D063065:Organophosphonates / Q000276:immunology / Q000378:metabolism; D017386:Sequence Homology, Amino Acid
D017151:Antibodies, Catalytic; D000911:Antibodies, Monoclonal; D004952:Esters; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007135:Immunoglobulin Variable Region; D009596:Nitrophenols; D063065:Organophosphonates; C085778:4-nitrophenyl phosphonate; D004247:DNA