Evidence map›Paper›PMID 19107192›Full record

ArticlePloS one2008

A simple detection method for low-affinity membrane protein interactions by baculoviral display.

Toshiko Sakihama, Takato Sato, Hiroko Iwanari, Toshio Kitamura, Shimon Sakaguchi, Tatsuhiko Kodama, Takao Hamakubo

Abstract readEvaluation Study
In one paragraph

Article in PloS one, 2008. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 3 papers.

0numbers the graph read from it
0cells of the map it votes in
3citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

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2 · The registry

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3 · Its place in the literature

Who cites it

3 citing papers in PubMed.

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4 · The record

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5 · Who and what money

Authors and funding

7 authors.

Toshiko SakihamaDepartment of Molecular Biology and Medicine, The University of Tokyo, Tokyo, Japan. sakihama@med.rcast.u-tokyo.ac.jp
Takato Sato
Hiroko Iwanari
Toshio Kitamura
Shimon Sakaguchi
Tatsuhiko Kodama
Takao Hamakubo

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

backgroundMembrane 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

findingsWe 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.

conclusionsWe 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.

Indexed as

Peptide LibraryAnimalsAntigens, SurfaceBaculoviridaeCD2 AntigensCD40 LigandCD58 AntigensCells, CulturedCloning, MolecularEnzyme-Linked Immunosorbent AssayHumansMembrane ProteinsMiceModels, BiologicalProtein BindingSpodopteraAntigens, SurfaceCD2 AntigensCD40 LigandCD58 AntigensMembrane ProteinsPeptide Library

Identifiers

PMID19107192
PMCPMC2602974

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.