Mechanistically different catalytic antibodies obtained from immunization with a single transition-state analog

Jincan Guo, Wei Huang, G. Wayne Zhou, Robert J. Fletterick, Thomas (Tom) Scanlan

Research output: Contribution to journalArticle

39 Citations (Scopus)

Abstract

The variable-region peptide sequence and steady-state kinetic behavior are compared for a family of catalytic antibodies that arose from the same immune response to a transition-state analog. The crystal structure of the most catalytically active member of the family (17E8) has been solved to 2.5 Å resolution and shows that the antibody active site contains a Ser(H99)- His(H35) (H = heavy chain) catalytic dyad analogous to the Ser-His-Asp catalytic triad of serine proteases. The variable-region peptide sequence of the next most active antibody (29G11) differs from that of 17E8 by nine heavy-chain point mutations, and results from computer modeling suggest that the three-dimensional structure of 29G11 is similar to that of 17E8. In addition, 29G11 is an efficient catalytic antibody; it possesses 26% of the hydrolytic activity of 17E8. There is one active-site mutation in 29G11 compared to 17E8; position 99 of the heavy chain of 29G11 contains a glycine residue in place of the nucleophilic serine at this position in 17E8. Consistent with this mutation, results from pH-rate studies and hydroxylamine partitioning experiments indicate that in contrast to the catalytic mechanism of 17E8, the mechanism of 29G11-catalyzed esterolysis does not feature nucleophilic catalysis.

Original languageEnglish (US)
Pages (from-to)1694-1698
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume92
Issue number5
DOIs
StatePublished - Feb 28 1995
Externally publishedYes

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Catalytic Antibodies
Immunization
Catalytic Domain
Peptides
Mutation
Hydroxylamine
Antibodies
Serine Proteases
Catalysis
Point Mutation
Glycine
Serine

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Mechanistically different catalytic antibodies obtained from immunization with a single transition-state analog. / Guo, Jincan; Huang, Wei; Zhou, G. Wayne; Fletterick, Robert J.; Scanlan, Thomas (Tom).

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 92, No. 5, 28.02.1995, p. 1694-1698.

Research output: Contribution to journalArticle

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