Mapping of the binding sites of human histamine N-methyltransferase (HNMT) monoclonal antibodies.

ABSTRACT:

Objective

Recently, we characterized mouse monoclonal antibodies that allow the specific and sensitive detection of human histamine N-methyltransferase (HNMT). To understand differences in binding characteristics and recognition of enzyme variants we mapped the antibody binding sites.

Methods

Fragments of human HNMT were expressed as glutathione S-transferase fusion proteins that were used for testing antibody binding on immunoblots. Combined information from species cross-reactivity, sequence comparison, protein structure, and binding site prediction software were used to localize the epitope recognized by each antibody.

Results

All eight monoclonal HNMT antibodies bound to linear epitopes in the C-terminal domain of the 292 amino acid protein. Of the five antibodies

SUBMITTER: Schwelberger HG 

PROVIDER: S-EPMC5633628 | biostudies-literature | 2017 Nov

REPOSITORIES: biostudies-literature