Abstract
The neuropeptide neurotensin (NT) interacts with two types of human receptors (hNTR) termed hNTR-1 and hNTR-2. This study describes a monoclonal antibody (MAb) specific for hNTR-1, B-N6. This MAb binds specifically to hNTR-1, but not to hNTR-2 transfected CHO cells. B-N6 and NT display a reciprocal competition and react in a similar way to trypsin, suggesting that the B-N6 epitope is at or close to the NT binding site on the third extracellular loop. Unlike B-N6, NT induces hNTR-1 internalization. Although neither NT-FITC nor B-N6 binding was detected by flow cytometry on different human cells, specific mRNA expression for hNTR-1 was detected in these cells. In CHO cells expressing hNTR-1 and a luciferase gene coupled to the krox24 reporter, B-N6 and the antagonist SR 48692 inhibited NT-induced intracellular activation of krox24 in a dose-dependent manner. From these results it is concluded that B-N6 is an antagonistic anti-hNTR-1 MAb.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Antibodies, Monoclonal / metabolism
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Antibodies, Monoclonal / pharmacology*
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Antibody Specificity
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Binding Sites, Antibody
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CHO Cells
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Cell Line
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Cricetinae
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DNA-Binding Proteins / genetics
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Early Growth Response Protein 1
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Flow Cytometry
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Humans
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Immediate-Early Proteins*
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Luciferases / genetics
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Mice
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Mice, Inbred BALB C
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Neurotensin / metabolism
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Neurotensin / pharmacology
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RNA, Messenger / analysis
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Receptors, Neurotensin / antagonists & inhibitors*
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Receptors, Neurotensin / genetics
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Receptors, Neurotensin / metabolism
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Recombinant Proteins
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Transcription Factors / genetics
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Transfection
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Trypsin / metabolism
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Tumor Cells, Cultured
Substances
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Antibodies, Monoclonal
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DNA-Binding Proteins
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EGR1 protein, human
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Early Growth Response Protein 1
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Egr1 protein, mouse
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Immediate-Early Proteins
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RNA, Messenger
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Receptors, Neurotensin
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Recombinant Proteins
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Transcription Factors
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Neurotensin
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Luciferases
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Trypsin