Effect Of LPS On Human CIAS1 Gene Expression And Structure And Function Analysis Of Cryopyrin

The innate immune system is in the first line of defence against microbial infection and cancer. The microbial sensing proteins, pattern-recognition receptors involved in innate immunity recognize pathogen-associated molecular patterns (PAMPs), which structural components often conserved of microorganisms. The CARTERPILLER (CARD, transcription enhancer, R/purine-binding, pyrin, lots leucine repeats) proteins, might be involved in intracellular recognition of microbes and their products, and induce inflammatory responses. The dysregulation of these processes due to mutations in the genes encoding these proteins is involved in numerous autoinflammatory disorders. Cryopyrin also known as NALP3, CIAS1 or PYPAF1, which encoded by the CIAS1 (cold-induced autoinflammatory syndrome 1) gene, is a CARTERPILLER's NALPs subfamily protein. Mutations within the CIAS1 gene are responsible for three autoinflammatory disorders: Muckle-Wells syndrome, familial cold autoinflammatory syndrome, and CINCA/ NOMID. Because of containing both PYD domain and NACHT+LRR domains, we believed that cryopyrin, which is responsible for some autoinflammatory diseases, is an important plasmosin involved in the intracellular recognition in innate immune system. However, little is known how cryopyrin is exactly "turned on" in living cells and how it stimulates the immune responses. In order to investigate the role of cryopyrin in the inflammatory diseases, and the function of each domain in cryopyrin in the intracellular recognition of PAMPs and in the cryopyrin-mediated signaling pathway, the methods of molecular biology and bioinformatics were used in this study.RT-PCR was used to detect the expression of CIAS1 gene in normal human leucocytes in the presence of LPS. On the other hand, structure-function analysis of cryopyrin based on available biological databases and bioinformatics prediction tools was performed for further insights into its molecule mechanisms. Several protein tools including 3D-PSSM, SPDBV, Rasmol, WHAT IF, GRAMM and ExPASy, were intergrated to model the three dimensional structure of three domains of cryopyrin,...