title: Crystallization and preliminary X-ray structural studies of a Melan-A pMHC-TCR complex.
abstract: Melanocytes are specialized pigmented cells that are found in all healthy skin tissue. In certain individuals, diseased melanocytes can form malignant tumours, melanomas, which cause the majority of skin-cancer-related deaths. The melanoma-associated antigenic peptides are presented on cell surfaces via the class I major histocompatibility complex (MHC). Among the melanoma-associated antigens, the melanoma self-antigen A/melanoma antigen recognized by T cells (Melan-A/MART-1) has attracted attention because of its wide expression in primary and metastatic melanomas. Here, a preliminary X-ray crystal structural study of a soluble cognate T-cell receptor (TCR) in complex with a pMHC presenting the Melan-A peptide (ELAGIGILTV) is reported. The TCR and pMHC were refolded, purified and mixed together to form complexes, which were crystallized using the sitting-drop vapour-diffusion method. Single TCR-pMHC complex crystals were cryocooled and used for data collection. Diffraction data showed that these crystals belonged to space group P4(1)/P4(3), with unit-cell parameters a = b = 120.4, c = 81.6 A. A complete data set was collected to 3.1 A and the structure is currently being analysed.
authors.affiliation: Nuffield Department of Clinical Medicine, John Radcliffe Hospital, Oxford University, Oxford OX3 9DU, England.
authors.forename: Fang
authors.initials: F
authors.lastname: Yuan
authors.forename: Theonie
authors.initials: T
authors.lastname: Georgiou
authors.forename: Theresa
authors.initials: T
authors.lastname: Hillon
authors.forename: Emma
authors.initials: E
authors.lastname: Gostick
authors.forename: David A
authors.initials: DA
authors.lastname: Price
authors.forename: Andrew K
authors.initials: AK
authors.lastname: Sewell
authors.forename: Ruth
authors.initials: R
authors.lastname: Moysey
authors.forename: Jessie
authors.initials: J
authors.lastname: Gavarret
authors.forename: Annelise
authors.initials: A
authors.lastname: Vuidepot
authors.forename: Malkit
authors.initials: M
authors.lastname: Sami
authors.forename: John I
authors.initials: JI
authors.lastname: Bell
authors.forename: George F
authors.initials: GF
authors.lastname: Gao
authors.forename: Pierre J
authors.initials: PJ
authors.lastname: Rizkallah
authors.forename: Bent K
authors.initials: BK
authors.lastname: Jakobsen
mesh_terms: D000951:Antigens, Neoplasm / Q000737:chemistry* / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D003460:Crystallization; D018076:DNA, Complementary; D006801:Humans; D058965:MART-1 Antigen; D008285:Major Histocompatibility Complex; D008544:Melanocytes / Q000502:physiology; D008545:Melanoma / Q000737:chemistry; D009363:Neoplasm Proteins / Q000737:chemistry* / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D010957:Plasmids; D011948:Receptors, Antigen, T-Cell / Q000276:immunology; D011994:Recombinant Proteins / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification; D012878:Skin Neoplasms / Q000737:chemistry; D014961:X-Ray Diffraction
chemical_list: D000951:Antigens, Neoplasm; D018076:DNA, Complementary; D058965:MART-1 Antigen; C548667:MLANA protein, human; D009363:Neoplasm Proteins; D011948:Receptors, Antigen, T-Cell; D011994:Recombinant Proteins