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Sequence Similarity and Cross‐Reactivity of a Diels–Alder Catalyst and an Anti‐Progesterone Antibody
Author(s) -
Haynes Matthew R.,
Lenz Martin,
Taussig Michael J.,
Wilson Ian A.,
Hilvert Donald
Publication year - 1996
Publication title -
israel journal of chemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.908
H-Index - 54
eISSN - 1869-5868
pISSN - 0021-2148
DOI - 10.1002/ijch.199600021
Subject(s) - chemistry , catalysis , mutagenesis , complementarity determining region , van der waals force , stereochemistry , active site , antibody , site directed mutagenesis , hydrogen bond , complementarity (molecular biology) , gene , peptide sequence , combinatorial chemistry , biochemistry , molecule , mutation , organic chemistry , genetics , biology , mutant
Antibody 1E9 catalyzes the bimolecular Diels–Alder reaction between tetrachlorothiophene dioxide and N ‐ethylmaleimide with high catalytic efficiency. The genes encoding the heavy and light chains were cloned, sequenced, and expressed as a mouse/human chimera in a murine hybridoma. Comparison with the closely related and structurally characterized anti‐progesterone antibody DB3 has permitted identification of the residues that are likely to line the binding cavity. In addition to confirming the expectation of a highly hydrophobic active site, modeling results suggest that shape complementarity, achieved mainly through van der Waals rather than hydrogen bonding or electrostatic interactions, is likely to be the dominant factor in achieving effective preorganization of the substrates for catalysis. These conclusions can now be investigated through targeted mutagenesis of the putative active site residues exploiting the mammalian expression system.