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. 2014 May 28;289(28):19799–19809. doi: 10.1074/jbc.M113.544627

FIGURE 4.

FIGURE 4.

The influence of swapping the POTRA domains of TpsB1 and TpsB2 on the secretion of TPS2a and TPS1 constructs and full-length TpsA1 and TpsA2 proteins. A, immunoblots of whole cell lysates (C) and culture supernatants (S) of N. meningitidis HB-1 tpsB1::kan/tpsB2::gen cells carrying plasmids encoding a wild-type or mutated TpsB as indicated above the lanes in combination with the TPS1 or TPS2a construct. B, immunoblots of whole cell lysates and culture supernatants of N. meningitidis HB-1 tpsB1::kan/tpsB2::gen cells carrying plasmids encoding a wild-type or mutated TpsB without TPS construct to analyze secretion of full-length TpsAs. The cells were grown in the presence (+) or absence (−) of 0.01 mm IPTG for expression of the TpsB. The blots were incubated with antisera against a TpsB or a TPS domain, as indicated on the right. On the left, the molecular weight markers are indicated. The full-length TpsA-derived bands in B are indicated by closed arrowheads (∼240, 200, and 75 kDa for TpsA1 and ∼250 and 260 kDa for TpsA2, respectively). A distinct background band detected by the TPS1 antiserum is indicated by an open arrowhead (19).