Fig 4. PamA K208(pBPa), PamA K278(pBPa), PamA K314(pBPa), PamA K382(pBPa) and PamA K395(pBPa) proteins crosslinked to unidentified endogenous proteins in N. meningitidis.

UV crosslinking of Pam K208amb, K278amb, K309amb, K341amb, K395amb, and K382amb mutants expressed by pyrrolysine-based amber suppression with pBPa in ΔpamA N. meningitidis. Bacteria that grew on one-fourth of a GC agar plate in the presence of 1 mM pBPa were crosslinked by UV light irradiation for 2 hours and then the PamA K(pBPa)-His₆ protein was purified. An aliquot was analyzed by anti-His₆ mAb. The black arrow shows the full-length PamA K-amb protein expressed by pyrrolysine-based amber suppression with pBPa, and the gray arrow indicates protein complexes crosslinked between PamA K(pBPa) and unidentified endogenous proteins in N. meningitidis.