The iron-regulated protein FrpD from Neisseria meningitidis is an outer membrane lipoprotein that interacts with very high affinity (Kd ~ 0.2 nM) with the N-terminal domain of FrpC, a Type I-secreted protein from the Repeat in ToXin (RTX) protein family. In the presence of Ca2+, FrpC undergoes Ca2+ -dependent protein trans-splicing that includes an autocatalytic cleavage of the Asp414-Pro415 peptide bond and formation of an Asp414-Lys isopeptide bond. Here, we report the high-resolution structure of FrpD and describe the structure-function relationships underlying the interaction between FrpD and FrpC1-414. We identified FrpD residues involved in FrpC1-414 binding, which enabled localization of FrpD within the low-resolution SAXS model of the FrpD-FrpC1-414 complex. Moreover, the trans-splicing activity of FrpC resulted in covalent linkage of the FrpC1-414 fragment to plasma membrane proteins of epithelial cells in vitro, suggesting that formation of the FrpD-FrpC1-414 complex may be involved in the interaction of meningococci with the host cell surface.

Center for Nanobiology and Structural Biology Institute of Microbiology Czech Academy of Sciences Zamek 136 37333 Nove Hrady Czech Republic

EMBL Hamburg Outstation c o DESY Notkestrasse 85 D 22603 Hamburg Germany

Faculty of Science University of South Bohemia Ceske Budejovice Branisovska 1760 37005 Ceske Budejovice Czech Republic

Institute of Microbiology Czech Academy of Sciences Videnska 1083 14220 Prague Czech Republic

Institute of Organic Chemistry and Biochemistry Czech Academy of Sciences Flemingovo nam 2 16610 Prague Czech Republic

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Structural Basis of Ca2+-Dependent Self-Processing Activity of Repeat-in-Toxin Proteins