cDNA cloning and expression of a hamster alpha-thrombin receptor coupled to Ca2+ mobilization

FEBS Lett. 1991 Aug 19;288(1-2):123-8. doi: 10.1016/0014-5793(91)81017-3.


The serine protease alpha-thrombin (thrombin) potently stimulates G-protein-coupled signaling pathways and DNA synthesis in CCL39 hamster lung fibroblasts. To clone a thrombin receptor cDNA, selective amplification of mRNA sequences displaying homology to the transmembrane domains of G-protein-coupled receptor genes was performed by polymerase chain reaction. Using reverse transcribed poly(A)+ RNA from CCL39 cells and degenerate primers corresponding to conserved regions of several phospholipase C-coupled receptors, three novel putative receptor sequences were identified. One corresponds to an mRNA transcript of 3.4 kb in CCL39 cells and a relatively abundant cDNA. Microinjection of RNA transcribed in vitro from this cDNA in Xenopus oocytes leads to the expression of a functional thrombin receptor. The hamster thrombin receptor consists of 427 amino acid residues with 8 hydrophobic domains, including one at the extreme N-terminus that is likely to represent a signal peptide. A thrombin consensus cleavage site is present in the N-terminal extracellular region of the receptor sequence followed by a negatively charged cluster of residues present in a number of proteins that interact with the anion-binding exosite of thrombin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Blotting, Northern
  • Calcium / metabolism*
  • Cloning, Molecular
  • Cricetinae
  • Cricetulus
  • GTP-Binding Proteins / genetics*
  • Humans
  • Molecular Sequence Data
  • Oocytes / metabolism
  • Polymerase Chain Reaction
  • Receptors, Cell Surface / genetics*
  • Receptors, Thrombin
  • Recombinant Proteins / biosynthesis
  • Sequence Alignment
  • Type C Phospholipases / genetics*
  • Xenopus


  • Receptors, Cell Surface
  • Receptors, Thrombin
  • Recombinant Proteins
  • Type C Phospholipases
  • GTP-Binding Proteins
  • Calcium

Associated data

  • GENBANK/M80612
  • GENBANK/S53371
  • GENBANK/X58797
  • GENBANK/X58798
  • GENBANK/X58799
  • GENBANK/X58800
  • GENBANK/X58801
  • GENBANK/X58802
  • GENBANK/X58803
  • GENBANK/X61958