FIG. 3.
SDS-PAGE of the gingipain complex and the corresponding patient sera reactivity in Western blots. (a) N-terminal sequencing by automated Edman degradation identified the composition of the bands in the denaturing (boiled and reduced) SDS-PAGE preparation of gingipain as RgpA (i), Kgpcat (ii), RgpAcat plus HA1 (iii), Kgp39 (iv), truncated HA1 + HA4 (v), and truncated RgpAcat (vi), HA2 (vii), and HA3 (viii). (b to d) Western blots of whole sera from disease patients against the gingipain complex (b), sera predepleted of anti-P. gingivalis LPS antibodies (c), and sera against the deglycosylated gingipain complex (d). 2B2 and 5A1 denote MAbs recognizing HA1, HA3, and Kgp39 and HA1, HA2, and Kgp39, respectively. Disease patients are numbered in the bar below panel b; number-prime values denote posttreatment serum from the same numbered patient. The āCā denotes serum from one representative control subject. Right arrows represent molecular weight markers: 118, 82, 50.4, 33.4, 26.7, and 19.6 kDa, respectively.