[ODP-146/W9-7] Examination of a meningococcal function-unknown protein with unnatural amino acid photocrosslinkers
Our previous genetic analyses revealed that the hypothetical protein, NMB1345, plays a crucial role in meningococcal infection in human brain microvascular endothelial cells; however, NMB1345 has no homology to any identified protein in databases and its physiological function could not be elucidated using pre-existing methods. In the present study, we examined the biological function(s) of the NMB1345 protein by using ncAAs-encoded photocrosslinking probes into N. meningitidis. The results revealed that the NMB1345 protein directly interacts with PilE, a major component of meningococcal pili, and further physicochemical and genetic analyses showed that the interaction between the NMB1345 protein and PilE was important for both functional pilus formation and meningococcal infectious ability in N. meningitidis. The present study using this new methodology for N. meningitidis provides novel insights into meningococcal pathogenesis by assigning the function of a hypothetical protein. Moreover, our established UNAA incorporation system will facilitate broad range of studies for characterize function of proteins function in N. meningitidis as well as other pathogenic bacteria.