A Simple Detection Method for Low-affinity Membrane Protein Interactions by Baculoviral Display

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2008 Dec 25

PMID 19107192

Citations 3

Authors

Toshiko Sakihama

Takato Sato

Hiroko Iwanari

Toshio Kitamura

Shimon Sakaguchi

Tatsuhiko Kodama

Takao Hamakubo

Affiliations

Soon will be listed here.

Abstract

Background: Membrane protein interactions play an important role in cell-to-cell recognition in various biological activities such as in the immune or neural system. Nevertheless, there has remained the major obstacle of expression of the membrane proteins in their active form. Recently, we and other investigators found that functional membrane proteins express on baculovirus particles (budded virus, BV). In this study, we applied this BV display system to detect interaction between membrane proteins important for cell-to-cell interaction in immune system.

Methodology/principal Findings: We infected Sf9 cells with recombinant baculovirus encoding the T cell membrane protein CD2 or its ligand CD58 and recovered the BV. We detected specific interaction between CD2-displaying BV and CD58-displaying BV by an enzyme-linked immunosorbent assay (ELISA). Using this system, we also detected specific interaction between two other membrane receptor-ligand pairs, CD40-CD40 ligand (CD40L), and glucocorticoid-induced TNFR family-related protein (GITR)-GITR ligand (GITRL). Furthermore, we observed specific binding of BV displaying CD58, CD40L, or GITRL to cells naturally expressing their respective receptors by flowcytometric analysis using anti-baculoviral gp64 antibody. Finally we isolated CD2 cDNA from a cDNA expression library by magnetic separation using CD58-displaying BV and anti-gp64 antibody.

Conclusions: We found the BV display system worked effectively in the detection of the interaction of membrane proteins. Since various membrane proteins and their oligomeric complexes can be displayed on BV in the native form, this BV display system should prove highly useful in the search for natural ligands or to develop screening systems for therapeutic antibodies and/or compounds.

Citing Articles

Improving T cell responses to modified peptides in tumor vaccines.

Buhrman J, Slansky J Immunol Res. 2012; 55(1-3):34-47.

PMID: 22936035 PMC: 3952016. DOI: 10.1007/s12026-012-8348-9.

Purification of functional baculovirus particles from silkworm larval hemolymph and their use as nanoparticles for the detection of human prorenin receptor (PRR) binding.

Kato T, Suzuki F, Park E BMC Biotechnol. 2011; 11:60.

PMID: 21635720 PMC: 3118113. DOI: 10.1186/1472-6750-11-60.

A novel screening system for claudin binder using baculoviral display.

Kakutani H, Takahashi A, Kondoh M, Saito Y, Yamaura T, Sakihama T PLoS One. 2011; 6(2):e16611.

PMID: 21339813 PMC: 3038848. DOI: 10.1371/journal.pone.0016611.

References

Hayashi I, Urano Y, Fukuda R, Isoo N, Kodama T, Hamakubo T . Selective reconstitution and recovery of functional gamma-secretase complex on budded baculovirus particles. J Biol Chem. 2004; 279(36):38040-6. DOI: 10.1074/jbc.M405597200. View

Saitoh R, Ohtomo T, Ito Y, Nezu J, Kimura N, Funahashi S . Recovery of functional peptide transporter PepT1 in budded baculovirus fraction. Protein Expr Purif. 2005; 46(1):130-5. DOI: 10.1016/j.pep.2005.08.008. View

Arulanandam A, Moingeon P, Concino M, Recny M, Kato K, Yagita H . A soluble multimeric recombinant CD2 protein identifies CD48 as a low affinity ligand for human CD2: divergence of CD2 ligands during the evolution of humans and mice. J Exp Med. 1993; 177(5):1439-50. PMC: 2191021. DOI: 10.1084/jem.177.5.1439. View

Morita S, Kojima T, Kitamura T . Plat-E: an efficient and stable system for transient packaging of retroviruses. Gene Ther. 2000; 7(12):1063-6. DOI: 10.1038/sj.gt.3301206. View

Liu J, Chow V, Jois S . A novel, rapid and sensitive heterotypic cell adhesion assay for CD2-CD58 interaction, and its application for testing inhibitory peptides. J Immunol Methods. 2004; 291(1-2):39-49. DOI: 10.1016/j.jim.2004.04.026. View

Abad L, Neumann M, Tobias L, Obenauer-Kutner L, Jacobs S, Cullen C . Development of a biosensor-based method for detection and isotyping of antibody responses to adenoviral-based gene therapy vectors. Anal Biochem. 2002; 310(1):107-13. DOI: 10.1016/s0003-2697(02)00314-7. View

Shiratori I, Ogasawara K, Saito T, Lanier L, Arase H . Activation of natural killer cells and dendritic cells upon recognition of a novel CD99-like ligand by paired immunoglobulin-like type 2 receptor. J Exp Med. 2004; 199(4):525-33. PMC: 2211832. DOI: 10.1084/jem.20031885. View

Sakihama T, Masuda K, Sato T, Doi T, Kodama T, Hamakubo T . Functional reconstitution of G-protein-coupled receptor-mediated adenylyl cyclase activation by a baculoviral co-display system. J Biotechnol. 2008; 135(1):28-33. DOI: 10.1016/j.jbiotec.2008.02.022. View

Watts T . TNF/TNFR family members in costimulation of T cell responses. Annu Rev Immunol. 2005; 23:23-68. DOI: 10.1146/annurev.immunol.23.021704.115839. View

10.

Brown M, Shaw S . T-cell activation: interplay at the interface. Curr Biol. 1999; 9(1):R26-8. DOI: 10.1016/s0960-9822(99)80038-4. View

11.

Clayton L, Sieh M, PIOUS D, Reinherz E . Identification of human CD4 residues affecting class II MHC versus HIV-1 gp120 binding. Nature. 1989; 339(6225):548-51. DOI: 10.1038/339548a0. View

12.

Suda T, Takahashi T, Golstein P, Nagata S . Molecular cloning and expression of the Fas ligand, a novel member of the tumor necrosis factor family. Cell. 1993; 75(6):1169-78. DOI: 10.1016/0092-8674(93)90326-l. View

13.

Van Broekhoven C, Altin J . A novel system for convenient detection of low-affinity receptor-ligand interactions: chelator-lipid liposomes engrafted with recombinant CD4 bind to cells expressing MHC class II. Immunol Cell Biol. 2001; 79(3):274-84. DOI: 10.1046/j.1440-1711.2001.01010.x. View

14.

Arulanandam A, Withka J, Wyss D, Wagner G, Kister A, Pallai P . The CD58 (LFA-3) binding site is a localized and highly charged surface area on the AGFCC'C" face of the human CD2 adhesion domain. Proc Natl Acad Sci U S A. 1993; 90(24):11613-7. PMC: 48034. DOI: 10.1073/pnas.90.24.11613. View

15.

Schonbeck U, Libby P . The CD40/CD154 receptor/ligand dyad. Cell Mol Life Sci. 2001; 58(1):4-43. PMC: 11146501. DOI: 10.1007/pl00000776. View

16.

Masuda K, Itoh H, Sakihama T, Akiyama C, Takahashi K, Fukuda R . A combinatorial G protein-coupled receptor reconstitution system on budded baculovirus. Evidence for Galpha and Galphao coupling to a human leukotriene B4 receptor. J Biol Chem. 2003; 278(27):24552-62. DOI: 10.1074/jbc.M302801200. View

17.

Davis S, Ikemizu S, Wild M, van der Merwe P . CD2 and the nature of protein interactions mediating cell-cell recognition. Immunol Rev. 1998; 163:217-36. DOI: 10.1111/j.1600-065x.1998.tb01199.x. View

18.

Loisel T, Ansanay H, Gay B, BOULANGER P, Strosberg A, Marullo S . Recovery of homogeneous and functional beta 2-adrenergic receptors from extracellular baculovirus particles. Nat Biotechnol. 1997; 15(12):1300-4. DOI: 10.1038/nbt1197-1300. View

19.

Shimizu J, Yamazaki S, Takahashi T, Ishida Y, Sakaguchi S . Stimulation of CD25(+)CD4(+) regulatory T cells through GITR breaks immunological self-tolerance. Nat Immunol. 2002; 3(2):135-42. DOI: 10.1038/ni759. View

20.

Hoffman T, Canziani G, Jia L, Rucker J, Doms R . A biosensor assay for studying ligand-membrane receptor interactions: binding of antibodies and HIV-1 Env to chemokine receptors. Proc Natl Acad Sci U S A. 2000; 97(21):11215-20. PMC: 17180. DOI: 10.1073/pnas.190274097. View