Poster Presentation The 44th Lorne Conference on Protein Structure and Function 2019

RECOMBINANT EXPRESSION OF A PROBLEMATIC PROTEIN (#107)

Larissa Doughty 1 2 , Craig Morton 2 , Michael Parker 1 2
  1. St. Vincent's Institute of Medical Research, Fitzroy, VIC, Australia
  2. Biochemistry and Molecular Biology, Bio21 Molecular Science and Biotechnology Institute, University of Melbourne, Parkville, Victoria, Australia

CD151 is a member of the tetraspanin family of transmembrane proteins, expressed in almost all cell types and tissues.[1, 2] The large extracellular loop (LEL) of CD151 is involved in protein-protein interactions that regulate cell adhesion, motility and proliferation and has been found to participate in nearly all stages of cancer progression.[3-5]  The LEL of CD151 exhibits high sequence variability from other tetraspanins and it has been shown that cancer metastasis is inhibited by anti-CD151 antibodies. This makes the LEL an attractive target for the development of inhibiting compounds that are selective for CD151.[6]

In spite of the numerous in vivo studies of CD151, structural information on the LEL is limited. After trialling many expression methods to produce recombinant LEL, I have been able to produce protein.  I present here conformational and stability studies of the monomeric protein.

 

  1. Sincock, P.M., G. Mayrhofer, and L.K. Ashman, Localization of the transmembrane 4 superfamily (TM4SF) member PETA-3 (CD151) in normal human tissues: comparison with CD9, CD63, and alpha5beta1 integrin. J Histochem Cytochem, 1997. 45(4): p. 515-25.
  2. Zoller, M., Tetraspanins: push and pull in suppressing and promoting metastasis. Nat Rev Cancer, 2009. 9(1): p. 40-55.
  3. Claas, C., et al., Association between the rat homologue of CO-029, a metastasis-associated tetraspanin molecule and consumption coagulopathy. J Cell Biol, 1998. 141(1): p. 267-80.
  4. Testa, J.E., et al., Eukaryotic expression cloning with an antimetastatic monoclonal antibody identifies a tetraspanin (PETA-3/CD151) as an effector of human tumor cell migration and metastasis. Cancer Res, 1999. 59(15): p. 3812-20.
  5. Kanetaka, K., et al., Overexpression of tetraspanin CO-029 in hepatocellular carcinoma. J Hepatol, 2001. 35(5): p. 637-42.
  6. Haeuw, J.F., et al., Tetraspanin CD151 as a target for antibody-based cancer immunotherapy. Biochem Soc Trans, 2011. 39(2): p. 553-8.