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Báo cáo khoa học: Soluble recombinant CD69 receptors optimized to have an exceptional physical and chemical stability display prolonged circulation and remain intact in the blood of mice

We investigated the soluble forms of the earliest activation antigen of human leukocyte CD69. This receptor is expressed at the cell surface as a type II homodimeric membrane protein. However, the elements necessary to prepare the soluble recombinant CD69 suitable for structural studies are a matter of controversy. | ỊFEBS Journal Soluble recombinant CD69 receptors optimized to have an exceptional physical and chemical stability display prolonged circulation and remain intact in the blood of mice Ondrej Vanek1 2 Monika Nalezkova3 Daniel Kavan1 2 Ivana Borovickova1 Petr Pompach1 2 Petr Novak2 Vinay Kumar2 Luca Vannucci2 Jiri Hudecek1 Katerina Hofbauerova2 4 Vladimir Kopeck Jr4 Jiri R t n 55 5 P t t l rJonl H6 155 PI hn55lolz6PQ ol 55 Hot 55 olz3 h moI ilz2 3 I 11Iz55Q t rr ilz3 I 11 Iz55 Q III Brynda Pen Koienko Jan Dohnaiek Pavel Kaderavek Josei Chmeiik Lukas xjoruik Lukas Zidek3 Vladimir Sklenar3 and Karel Bezouska1 2 1 Department of Biochemistry Faculty of Science Charles University Prague Czech Republic 2 Institute of Microbiology Academy of Sciences of Czech Republic Prague Czech Republic 3 NationalCentre for Biomolecular Research Faculty of Science Masaryk University Brno Czech Republic 4 Institute of Physics Faculty of Mathematics and Physics Charles University Prague Czech Republic 5 Institute of Molecular Genetics Academy of Sciences of Czech Republic Prague Czech Republic 6 Institute of Macromolecular Chemistry Academy of Sciences of Czech Republic Prague Czech Republic Keywords C-type lectin leukocyte activation plasma clearance refolding stability Correspondence K. Bezouska Department of Biochemistry Faculty of Science Charles University Prague Hlavova 8 CZ-12840 Praha 2 Czech Republic Fax 420 2 4172 1143 Tel 420 2 4106 2383 E-mail bezouska@biomed.cas.cz These authors contributed equally to this work Received 5 June 2008 revised 2 September 2008 accepted 11 September 2008 doi 10.1111 j.1742-4658.2008.06683.x We investigated the soluble forms of the earliest activation antigen of human leukocyte CD69. This receptor is expressed at the cell surface as a type II homodimeric membrane protein. However the elements necessary to prepare the soluble recombinant CD69 suitable for structural studies are a matter of controversy. We describe the physical biochemical and in vivo