Crystallization and functional analysis of a soluble deglycosylated form of the human costimulatory molecule B7-1.
The interactions of B7-1 with CD28 and CTLA-4 modulate the course of human immune responses, making B7-1 an important target for developing structure-based therapeutics. B7-1 is, however, one of the most heavily glycosylated proteins found at the leukocyte cell surface, complicating the structural a...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Journal article |
| Language: | English |
| Published: |
2001
|
| _version_ | 1797068718155497472 |
|---|---|
| author | Davis, S Ikemizu, S Collins, A Fennelly, J Harlos, K Jones, E Stuart, D |
| author_facet | Davis, S Ikemizu, S Collins, A Fennelly, J Harlos, K Jones, E Stuart, D |
| author_sort | Davis, S |
| collection | OXFORD |
| description | The interactions of B7-1 with CD28 and CTLA-4 modulate the course of human immune responses, making B7-1 an important target for developing structure-based therapeutics. B7-1 is, however, one of the most heavily glycosylated proteins found at the leukocyte cell surface, complicating the structural analysis of this molecule. Methods for the production, crystallization and selenomethionine labelling of a soluble deglycosylated form of this molecule are described. The protein readily forms both tetragonal plate and bipyramidal crystals belonging to space groups I4(1)22, with unit-cell parameters a = b = 56.9, c = 298.7 A, and P4(1)22 (or P4(3)22), with unit-cell parameters a = b = 89.0, c = 261.9 A, respectively. The I4(1)22 and primitive crystal forms diffract to 2.7 and 3.5 A, respectively. Surface plasmon resonance-based assays indicate that the ligand-binding properties of sB7-1 are unaffected by deglycosylation. Since none of the methods relied on any special structural properties of sB7-1, it is proposed that this novel combination of procedures could in principle be adapted to the systematic analysis of many other glycoproteins of structural or functional interest. |
| first_indexed | 2024-03-06T22:14:13Z |
| format | Journal article |
| id | oxford-uuid:52d8b01c-793f-41a4-9902-6b13ca5b6cfe |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-06T22:14:13Z |
| publishDate | 2001 |
| record_format | dspace |
| spelling | oxford-uuid:52d8b01c-793f-41a4-9902-6b13ca5b6cfe2022-03-26T16:27:56ZCrystallization and functional analysis of a soluble deglycosylated form of the human costimulatory molecule B7-1.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:52d8b01c-793f-41a4-9902-6b13ca5b6cfeEnglishSymplectic Elements at Oxford2001Davis, SIkemizu, SCollins, AFennelly, JHarlos, KJones, EStuart, DThe interactions of B7-1 with CD28 and CTLA-4 modulate the course of human immune responses, making B7-1 an important target for developing structure-based therapeutics. B7-1 is, however, one of the most heavily glycosylated proteins found at the leukocyte cell surface, complicating the structural analysis of this molecule. Methods for the production, crystallization and selenomethionine labelling of a soluble deglycosylated form of this molecule are described. The protein readily forms both tetragonal plate and bipyramidal crystals belonging to space groups I4(1)22, with unit-cell parameters a = b = 56.9, c = 298.7 A, and P4(1)22 (or P4(3)22), with unit-cell parameters a = b = 89.0, c = 261.9 A, respectively. The I4(1)22 and primitive crystal forms diffract to 2.7 and 3.5 A, respectively. Surface plasmon resonance-based assays indicate that the ligand-binding properties of sB7-1 are unaffected by deglycosylation. Since none of the methods relied on any special structural properties of sB7-1, it is proposed that this novel combination of procedures could in principle be adapted to the systematic analysis of many other glycoproteins of structural or functional interest. |
| spellingShingle | Davis, S Ikemizu, S Collins, A Fennelly, J Harlos, K Jones, E Stuart, D Crystallization and functional analysis of a soluble deglycosylated form of the human costimulatory molecule B7-1. |
| title | Crystallization and functional analysis of a soluble deglycosylated form of the human costimulatory molecule B7-1. |
| title_full | Crystallization and functional analysis of a soluble deglycosylated form of the human costimulatory molecule B7-1. |
| title_fullStr | Crystallization and functional analysis of a soluble deglycosylated form of the human costimulatory molecule B7-1. |
| title_full_unstemmed | Crystallization and functional analysis of a soluble deglycosylated form of the human costimulatory molecule B7-1. |
| title_short | Crystallization and functional analysis of a soluble deglycosylated form of the human costimulatory molecule B7-1. |
| title_sort | crystallization and functional analysis of a soluble deglycosylated form of the human costimulatory molecule b7 1 |
| work_keys_str_mv | AT daviss crystallizationandfunctionalanalysisofasolubledeglycosylatedformofthehumancostimulatorymoleculeb71 AT ikemizus crystallizationandfunctionalanalysisofasolubledeglycosylatedformofthehumancostimulatorymoleculeb71 AT collinsa crystallizationandfunctionalanalysisofasolubledeglycosylatedformofthehumancostimulatorymoleculeb71 AT fennellyj crystallizationandfunctionalanalysisofasolubledeglycosylatedformofthehumancostimulatorymoleculeb71 AT harlosk crystallizationandfunctionalanalysisofasolubledeglycosylatedformofthehumancostimulatorymoleculeb71 AT jonese crystallizationandfunctionalanalysisofasolubledeglycosylatedformofthehumancostimulatorymoleculeb71 AT stuartd crystallizationandfunctionalanalysisofasolubledeglycosylatedformofthehumancostimulatorymoleculeb71 |