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.
Laboratory of Protein Synthesis and Expression, Institute for Protein Research, Osaka University, Osaka, Japan.
Takeshi
T
Sangawa
Terukazu
T
Nogi
Junichi
J
Takagi
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