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Protein Membrane Overlay Assay: A Protocol to Test Interaction Between Soluble and Insoluble Proteins in vitro
Published on: August 14, 2011
Porcine CD83 is a glycosylated dimeric protein existing naturally in membrane-bound and soluble forms
Shanshan Huo1, Jianlou Zhang1, Fengyang Wu2
1Laboratory of Molecular Virology and Immunology, College of Animal Science and Technology/College of Veterinary Medicine, Agricultural University of Hebei, Baoding, Hebei, 071000, China; Hebei Engineering and Technology Research Center of Veterinary Biotechnology, Baoding, Hebei, 071000, China.
Abstract:
Human and mouse CD83 have been well characteized, however, the other mammalian CD83 genes have not been cloned and characterized. In this study, the porcine CD83 (pCD83) was cloned, expressed and characterized, and showed that the pCD83 gene has 81% and 74% homologies with humans and mice, respectively, which was identified to be glycosylated when expressed in eukaryotic cells, existing naturally in two forms: membrance-bound CD83 (mCD83) and soluble CD83 (sCD83), the latter was identified to be generated mainly from mCD83 by proteolytic shedding. The pCD83 was a dimmer mediated by intermolecular disulfide bond formed by the fifth cysteine in the exrtracellular domain. Functionally, the recombinant porcine sCD83 was preliminarily tested to have the ability to inhibit DC-mediated T cell activition. This study provided necessary fundation for further investigation on pCD83 functions.
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