[A structure-activity study of a catalytic antiidiotypic antibody to the human erythrocyte acetylcholinesterase].
The catalytic monoclonal antibody 9A8 (MA 9A8), antiidiotypic to the antibody AE-2 (MA AE2) produced to the active site of acetyl cholinesterase from human erythrocytes, was subjected to a structure-function study. The specific binding of MA 9A8 to MA AE2 (K 2.26 x 10(9) M-1) was shown by the method of surface plasmon resonance, and the functional activity of MA 9A8 was demonstrated. Unlike acetyl cholinesterase, this antibody specifically reacted with the irreversible phosphonate inhibitors of esterases. A peptide map of MA 9A8 was analyzed by MALDI mass spectrometry. The Ser99 residue of its heavy chain was shown to be within the active site of the catalytic antibody. A computer modeling of the MA 9A8 active site suggested the existence of a catalytic dyad formed by Ser99 and His35. A comparison of the tertiary structures of the MA 9A8 and the 17E8 monoclonal antibody, which also exhibited an esterase activity and was produced to the stable analogue of the reaction transition state, indicated a practically complete coincidence of the structures of their presumed active sites.
Aleksandrova
E S
ES
Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, GSP Moscow, 117997 Russia. lenik@ibch.ru
Koralevski
F
F
Titov
M I
MI
Demin
A V
AV
Kozyr'
A V
AV
Kolesnikov
A V
AV
Tramontano
A
A
Paul
S
S
Thomas
D
D
Gabibov
A G
AG
Gnuchev
N V
NV
Friboulet
A
A
D000110:Acetylcholinesterase / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism; D000595:Amino Acid Sequence; D000888:Antibodies, Anti-Idiotypic / Q000737:chemistry* / Q000378:metabolism*; D017151:Antibodies, Catalytic / Q000737:chemistry* / Q000378:metabolism*; D002800:Cholinesterase Inhibitors / Q000737:chemistry / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004912:Erythrocytes / Q000201:enzymology / Q000276:immunology*; D006639:Histidine; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011487:Protein Conformation; D012694:Serine; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D020349:Surface Plasmon Resonance
D000888:Antibodies, Anti-Idiotypic; D017151:Antibodies, Catalytic; D002800:Cholinesterase Inhibitors; D004791:Enzyme Inhibitors; D012694:Serine; D006639:Histidine; D000110:Acetylcholinesterase