Characterization of monoclonal antibodies directed against the rat neurotensin receptor NTS1.
G protein-coupled receptors (GPCRs) are integral membrane proteins that mediate cellular responses to a variety of ligands and represent major drug targets. Despite their medical importance, detailed structural information is limited because only one GPCR has been crystallized and its structure determined. To develop tools to aid in the formation of well-ordered crystals, we generated monoclonal antibodies with high affinity to the rat neurotensin receptor. All antibodies bound to the C-terminus of the receptor, which may reflect the selection strategy used to identify high-affinity binders. Further characterization revealed that some antibodies bound to the receptor in a sodium chloride sensitive manner, but others did not. Epitope mapping revealed distinct antigenic regions within the receptor C-terminus. Tight binding of Fab fragments to the receptor was verified by size exclusion chromatography.
Niebauer
Ronald T
RT
Department of Health and Human Services, Laboratory of Molecular Biology of the National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland, USA.
White
Jim F
JF
Fei
Zhizhong
Z
Grisshammer
Reinhard
R
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology; D001665:Binding Sites; D003460:Crystallization / Q000379:methods*; D018604:Epitope Mapping; D007140:Immunoglobulin Fab Fragments / Q000378:metabolism; D051381:Rats; D043562:Receptors, G-Protein-Coupled / Q000276:immunology; D018028:Receptors, Neurotensin / Q000276:immunology*; D012965:Sodium Chloride
D000911:Antibodies, Monoclonal; D007140:Immunoglobulin Fab Fragments; D043562:Receptors, G-Protein-Coupled; D018028:Receptors, Neurotensin; C114036:neurotensin type 1 receptor; D012965:Sodium Chloride