The SDS-PAGE containing connexin 43 is shown in Figure 5. There was a twofold increase in radioactivity compared with the supernatant, which was not different from background activity. In the next step of the further purification of the connexins, ie, of con-nexin 43, radioactivity was lost. Finally, connexin 43 was found, but no 14C-labelled AAP10 was detectable (Table 2)
Figure 5 SDS-Page of purified gap junctional membranes. Numbers indicate pufication steps. GB Gradient band, ie, the interface material of step 36; GP Gradient pellet; M Molecular weight marker. Lanes to the left of M were derivedfrom undiluted test volumes; lanes to the right of M were diluted 1:1 with buffer solution. Note the 43 kDa band containing connexin 43 from rabbit hearts. During further purification, connexin 43 was enriched in the gradient band
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TABLE 2 Radioactivity in pellets and supernatants following purification of connexin 43
|Pellet S21||83.30||Intracellular and membrane-associated proteins|
|Connexin 43||nd||Radiofluorography/PAGEgel electrophoresis|
nd None detectable. Numbers (Sx) refer to the step numbers given in the Methods section. Note that there was increased activity in the pellet of step 21