A murine monoclonal antibody that binds N-terminal extracellular segment of human protease-activated receptor-4.
Abstract A monoclonal antibody that recognizes native G protein coupled receptors (GPCR) is generally difficult to obtain. Protease-activated receptor-4 (PAR4) is a GPCR that plays an important role in platelet activation as a low-affinity thrombin receptor. By immunizing peptide corresponding to the N-terminal segment of human PAR4, we obtained a monoclonal antibody that recognizes cell surface expressed PAR4. Epitope mapping using a series of artificial fusion proteins that carry PAR4-derived peptide revealed that the recognition motif is fully contained within the 6-residue portion adjacent to the thrombin cleavage site. The antibody blocked PAR4 peptide cleavage by thrombin, suggesting its utility in the functional study of PAR4 signaling.
Sangawa
Takeshi
T
Laboratory of Protein Synthesis and Expression, Institute for Protein Research, Osaka University, Osaka, Japan.
Nogi
Terukazu
T
Takagi
Junichi
J
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D018604:Epitope Mapping; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D010455:Peptides; D017434:Protein Structure, Tertiary; D018179:Receptors, Thrombin / Q000737:chemistry / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000276:immunology; D013917:Thrombin / Q000378:metabolism
D000911:Antibodies, Monoclonal; D010455:Peptides; D018179:Receptors, Thrombin; D011993:Recombinant Fusion Proteins; D013917:Thrombin; C112908:protease-activated receptor 4