Multivalent recombinant proteins for probing functions of leucocyte surface proteins such as the CD200 receptor.
CD200 (OX2) is a membrane glycoprotein that interacts with a structurally related receptor (CD200R) involved in the regulation of macrophage function. The interaction is of low affinity (K(D) approximately 1 microm) but can be detected using CD200 displayed in a multivalent form on beads or with dim...
| Main Authors: | , , , , , |
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| Format: | Journal article |
| Language: | English |
| Published: |
2005
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| _version_ | 1797105337580388352 |
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| author | Voulgaraki, D Mitnacht-Kraus, R Letarte, M Foster-Cuevas, M Brown, M Barclay, N |
| author_facet | Voulgaraki, D Mitnacht-Kraus, R Letarte, M Foster-Cuevas, M Brown, M Barclay, N |
| author_sort | Voulgaraki, D |
| collection | OXFORD |
| description | CD200 (OX2) is a membrane glycoprotein that interacts with a structurally related receptor (CD200R) involved in the regulation of macrophage function. The interaction is of low affinity (K(D) approximately 1 microm) but can be detected using CD200 displayed in a multivalent form on beads or with dimeric fusion proteins consisting of the extracellular region of CD200 and immunoglobulin Fc regions. We prepared putative pentamers and trimers of mouse CD200 with sequences from cartilage oligomeric matrix protein (COMP) and surfactant protein D (SP-D), respectively. The COMP protein gave high-avidity binding and was a valuable tool for showing the interaction whilst the SP-D protein gave weak binding. In vivo experiments showed that an agonistic CD200R monoclonal antibody caused some amelioration in a model of experimental autoimmune encephalomyelitis but the COMP protein was cleared rapidly and had minimal effect. Pentameric constructs also allowed detection of the rat CD48/CD2 interaction, which is of much lower affinity (K(D) approximately 70 microm). These reagents may have an advantage over Fc-bearing hybrid molecules for probing cell surface proteins without side-effects due to the Fc regions. The CD200-COMP gave strong signals in protein microarrays, suggesting that such reagents may be valuable in high throughput detection of weak interactions. |
| first_indexed | 2024-03-07T06:46:04Z |
| format | Journal article |
| id | oxford-uuid:fae42ae7-c84e-4908-b695-893281bb1539 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T06:46:04Z |
| publishDate | 2005 |
| record_format | dspace |
| spelling | oxford-uuid:fae42ae7-c84e-4908-b695-893281bb15392022-03-27T13:09:43ZMultivalent recombinant proteins for probing functions of leucocyte surface proteins such as the CD200 receptor.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:fae42ae7-c84e-4908-b695-893281bb1539EnglishSymplectic Elements at Oxford2005Voulgaraki, DMitnacht-Kraus, RLetarte, MFoster-Cuevas, MBrown, MBarclay, NCD200 (OX2) is a membrane glycoprotein that interacts with a structurally related receptor (CD200R) involved in the regulation of macrophage function. The interaction is of low affinity (K(D) approximately 1 microm) but can be detected using CD200 displayed in a multivalent form on beads or with dimeric fusion proteins consisting of the extracellular region of CD200 and immunoglobulin Fc regions. We prepared putative pentamers and trimers of mouse CD200 with sequences from cartilage oligomeric matrix protein (COMP) and surfactant protein D (SP-D), respectively. The COMP protein gave high-avidity binding and was a valuable tool for showing the interaction whilst the SP-D protein gave weak binding. In vivo experiments showed that an agonistic CD200R monoclonal antibody caused some amelioration in a model of experimental autoimmune encephalomyelitis but the COMP protein was cleared rapidly and had minimal effect. Pentameric constructs also allowed detection of the rat CD48/CD2 interaction, which is of much lower affinity (K(D) approximately 70 microm). These reagents may have an advantage over Fc-bearing hybrid molecules for probing cell surface proteins without side-effects due to the Fc regions. The CD200-COMP gave strong signals in protein microarrays, suggesting that such reagents may be valuable in high throughput detection of weak interactions. |
| spellingShingle | Voulgaraki, D Mitnacht-Kraus, R Letarte, M Foster-Cuevas, M Brown, M Barclay, N Multivalent recombinant proteins for probing functions of leucocyte surface proteins such as the CD200 receptor. |
| title | Multivalent recombinant proteins for probing functions of leucocyte surface proteins such as the CD200 receptor. |
| title_full | Multivalent recombinant proteins for probing functions of leucocyte surface proteins such as the CD200 receptor. |
| title_fullStr | Multivalent recombinant proteins for probing functions of leucocyte surface proteins such as the CD200 receptor. |
| title_full_unstemmed | Multivalent recombinant proteins for probing functions of leucocyte surface proteins such as the CD200 receptor. |
| title_short | Multivalent recombinant proteins for probing functions of leucocyte surface proteins such as the CD200 receptor. |
| title_sort | multivalent recombinant proteins for probing functions of leucocyte surface proteins such as the cd200 receptor |
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