title: A soluble recombinant form of human leucocyte antigen-G 6 (srHLA-G6).
abstract: Human Leucocyte Antigen-G (HLA-G) is a non classical major histocompatibility complex (MHC) molecule that through RNA splicing can encode seven isoforms which are membrane bound (-G1, -G2, -G3 and -G4) and soluble (-G5, -G6 and -G7). HLA-G is described as important immune suppressor endogenous molecule to favor maternal-fetal tolerance, transplant survival and tumor immune scape. HLA-G shows low protein variability and a unique structural complexity that is related with the expression of different isoforms followed by biochemical processes, such as, proteolytic cleavage, molecular interactions, and protein ubiquitination. Studies with HLA-G have shown difficult to assess the role of the individual isoforms. Thus, the aim of this work was to obtain a HLA-G6 recombinant form. The results indicated the production of high homogeneous preparations of soluble recombinant HLA-G6 (srHLA-G6) with molecular mass 23,603.76 Da, determined by MALD-TOF/TOF. In addition, native and denatured srHLA-G6 were detected by ELISA, using commercial monoclonal antibodies. Finally, we developed a suitable methodology to express srHLA-G6 that could contribute in structural and functional studies involving specific isoforms.
authors.affiliation: Laboratório de Glicoimunologia, Universidade de São Paulo, Av. Café, s/n 14040-903, Ribeirão Preto, SP, Brazil.
authors.forename: Flávia Porto
authors.initials: FP
authors.lastname: Pelá
authors.affiliation: Laboratório de Cristalografia de Proteínas, Universidade de São Paulo, Av. Café, s/n 14040-903, Ribeirão Preto, SP, Brazil.
authors.forename: Joane Kathelen
authors.initials: JK
authors.lastname: Rustiguel
authors.affiliation: Laboratório de Glicoimunologia, Universidade de São Paulo, Av. Café, s/n 14040-903, Ribeirão Preto, SP, Brazil.
authors.forename: Lilian Cataldi
authors.initials: LC
authors.lastname: Rodrigues
authors.affiliation: Núcleo de Pesquisa em Produtos Naturais e Sintéticos, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Av. Café, s/n, 14040-903, Ribeirão Preto, SP, Brazil.
authors.forename: Jacqueline Nakau
authors.initials: JN
authors.lastname: Mendonça
authors.affiliation: Laboratório de Glicoimunologia, Universidade de São Paulo, Av. Café, s/n 14040-903, Ribeirão Preto, SP, Brazil.
authors.forename: Camillo Del Cistia
authors.initials: CDC
authors.lastname: Andrade
authors.affiliation: Núcleo de Pesquisa em Produtos Naturais e Sintéticos, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Av. Café, s/n, 14040-903, Ribeirão Preto, SP, Brazil.
authors.forename: Norberto Peporine
authors.initials: NP
authors.lastname: Lopes
authors.affiliation: Centro de Química de Proteínas, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Rua Tenente Catão Roxo, 2501, 14051-140, Ribeirão Preto, SP, Brazil.
authors.forename: José Cesar
authors.initials: JC
authors.lastname: Rosa
authors.affiliation: Laboratório de Cristalografia de Proteínas, Universidade de São Paulo, Av. Café, s/n 14040-903, Ribeirão Preto, SP, Brazil.
authors.forename: Maria Cristina
authors.initials: MC
authors.lastname: Nonato
authors.affiliation: CEA/DRF/IDMIT-Université Paris Sud-INSERM U1184/IMVA, 92265, Fontenay-aux-Roses, France.
authors.forename: Benoit
authors.initials: B
authors.lastname: Favier
authors.affiliation: Divisão de Imunologia Clínica, Departamento de Clínica Médica, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Av. Bandeirantes, 3900 - Monte Alegre, 14049-900, Ribeirão Preto, SP, Brazil.
authors.forename: Eduardo Antônio
authors.initials: EA
authors.lastname: Donadi
authors.affiliation: Laboratório de Glicoimunologia, Universidade de São Paulo, Av. Café, s/n 14040-903, Ribeirão Preto, SP, Brazil. Electronic address: mdbaruff@fcfrp.usp.br.
authors.forename: Marcelo
authors.initials: M
authors.lastname: Dias-Baruffi
mesh_terms: D001665:Binding Sites; D059951:HLA-G Antigens / Q000737:chemistry* / Q000276:immunology*; D006801:Humans; D008970:Molecular Weight; D011485:Protein Binding; D011994:Recombinant Proteins / Q000737:chemistry*; D012995:Solubility
chemical_list: D059951:HLA-G Antigens; D011994:Recombinant Proteins
keywords: Human leucocyte antigen-G6; MALD-TOF/TOF; MEM-G9; Recombinant protein expression